Download Methanol and Ethanol

Tracking the short-term fate of some organic pollutants of the
ISS Crew Latent Condensate (methanol, ethanol, and
isopropanol) when included in the nutrient solution for rocket
(Eruca sativa Mill.) hydroponic cultivation
Simona Proietti, Walter Stefanoni, Stefano Moscatello, Emanuele Pallozzi, Lucia
Grizzaffi, Ilaria Locantore, Carlo Calfapietra, Marco Lauteri, Alberto Battistelli.
CNR – IBAF Istituto di Biologia Agroalimentare e Forestale, Porano (TR) - Italy
Thales Alenia Space Italia Torino - Italy
DAFNE, Università della Tuscia, via San Camillo De Lellis snc, 01100 Viterbo (VT) - Italy
International Project:
Inflatable greenhouse for space plant food production and life support.
Italian Ministry of Foreign Affairs (MAE)
th
10 Review Conference on Scientific and Technological Cooperation Agreement between Italy and USA
Workshop on Bioregenerative Life Support Turin, 18-19 May 2015
During long term human space mission, water recovered from various streams should be
used as the basic component of nutrient solutions for plant cultivation.
For this end the quality of
recovery water needs to be
carefully evaluated since plants
growth and plant food quality
can be affected by the quality of
water used
5x10-3 %
8x10-4 %
4x10-4 %
Water streams from space
station can contains various
organic contaminants
Methanol, Ethanol, and
Iso-propanol (MEI) are
included in the list of
Wastewater Contaminants in
Crew Latent Condensate
(Wastewater Contaminants in Crew Latent Condensate: Handford, 2004).
Methanol, Ethanol, and Iso-propanol (MEI) are selected to investigate
the effects of relevant contaminants found in the ISS
on plant growth and on plant physiology.
Rocket plant (Eruca sativa Mill.)
grown from seeds in a growth chamber
for 4 weeks
in a floating system
Feeding of MEI were imposed on rocket seeds
and on rocket plants
With MEI
Do the rocket seeds germinate?
Methanol and isopropanol do not affect rocket
seeds germination also at high concentration (1%)
Ethanol affects seeds germination only at high
concentration (1% and 0,5%)
When the ethanol concentration was decreased
at 0,2% until the concentration found in the
water condensate of ISS the germination rate of
rocket seed is not affected.
8x10-4 % = ethanol
found in the
water condensate of ISS
Are there a rapid and
destructive effects on rocket
plants?
With MEI
Gas exchange measurements and fluorescence of chlorophyll a
using a Licor XT 6400 after feeding with 15mM of MEI
Feeding
Alcohols did not affected photosynthesis
rate nor fluorescence emission.
1,0
Fv/Fm
0,8
0,6
0,4
0,2
0,0
Control
Metanol
Ethanol
treatment
Isopropanol
Gas exchanges were unaltered by
alcohols also at higher concentration
to respect the concentration found in
the water streams of ISS.
Are they taken up and
accumulated by rocket plants?
MEI
Are they
accumulated?
Are they
translocated?
Leaf extracts from plants fed with
MEI 1% were analyzed by
Ion Chromatography ICS 5000
(Dionex ThermoFisher)
Are they
absorbed?
Methanol and Ethanol, but not isopropanol, were detected in leaves after feeding,
but a low concentration.
i2 f2
isoprop 0.07%
ED_1
ED_1
1 - isopropanolo - 7,400
1 - 17-2-2014 ma1 #78 [modified by Stefano]
30,0 2 - 17-2-2014 ma1 #88 [modified by Stefano, 1 peak manually assigned]
nC
25,0
20,0
15,0
Standard
Expected Isopropanol in leaf
10,0 1
5,0
2
-2,0
7,00
min
7,10
7,20
7,30
7,40
7,50
7,60
7,70
7,80
7,90
8,00
8,10
8,18
Methanol and Ethanol (and also Isopropanol, see
later) are taken up by the rocket root and
translocated to leaves, but do not accumulate to
high concentration.
What is the fate of MEI once they are inside the plant ?
CO2
(complete destruction)
Metabolism
Metabolites
Formaldehyde - Formic acid
Acetaldehyde - Acetic acid
Acetone
MEI in plant tissues
Accumulation (low)
Re-emission
in the atmosphere
Accumulation
Labeling experiments were carried out to test if MEI enter into the respiratory
metabolism using 13C enriched MEI
Feeding was carried out with a solution (15 mM)
of each alcohols containing a small proportion of
13C labeled molecules.
The presence of 13C was measured (d13C) on CO2
released by leaf respiration, by IRMS.
40
Controllo
20
0
Metanolo
24h
-20
-40
60
Rapp.
isotopico
Isotopic
ratio
40
20
Controllo
0
-20
-40
Etanolo
24h
60
d13C
Isotopic
ratio
Rapp.
isotopico
d13C
d13C
60
Isopropanol 15 mM
Ethanol 15 mM
Methanol 15 mM
Rapp.
isotopico
Isotopic
ratio
40
20
Controllo
0
-20
-40
Isopropanolo
24h
enriched MEI caused a remarkably change in d13C in the respiratory CO2
with a decreasing intensity with increasing alcohol molecular mass.
This clearly indicates the uptake of alcoholic compounds from plants and a fast turn-over of
labeled methanol and ethanol molecules as soon as they enter on the plant metabolism.
M and E can be metabolized to enter respiratory metabolism: plants can contribute to
degradation of M and E to CO2.
Feeding of
13C
Are their metabolic products
accumulated in plant tissues?
MEI
oxidation
oxidation
Formaldehyde
Methanol
Formic acid
CO2
oxidation
oxidation
Ethanol
oxidation
Acetaldehyde
Acetic acid
oxidation
Isopropanol
Acetone
Organic acids, as a product of alcohols
metabolism in plants were analyzed by
Dionex™ ICS-5000 from leaf extracts from
plants fed with MEI.
Accumulation of metabolic products of MEI in rocket leaves.
No accumulation of metabolic products of methanol
were detected by IC
1 - 24-02-2014B #13 [modified by Stefano]
2 - 24-02-2014B #22 [modified by Stefano]
3 - 24-02-2014B #24 [modified by Stefano]
e1 f2
e2 f1
c1 f2
CD_1
CD_1
CD_1
1 - Acetico
11,3
µS
Acetic acid in leaves
8,0
1 - 3,253
6,0
4,0
2,0
1
3
2
0,0
-2,4
2,961
min
3,050
3,100
3,150
3,200
3,250
3,300
3,350
3,400
3,450
3,500
3,550
3,616
An increment of acetic acid in leaves was detected after Ethanol feeding.
Are they released, in the atmosphere?
Are their metabolic producs released?
Methanol Ethanol
Isopropanol
oxidation
oxidation
Formaldehyde
Methanol
Formic acid
oxidation
oxidation
Ethanol
oxidation
Acetaldehyde
Acetic acid
oxidation
Isopropanol
Acetone
The emission in the atmosphere of MEI and relevant metabolites were
analyzed on line by Real-time trace gas analyzer solutions based on
Proton-transfer-reaction mass spectrometry (PTR-MS - Ionicon).
Methanol and formaldehyde and formic acid from methanol oxidation
Ethanol and acetaldehyde and acetic acid from ethanol oxidation
Isopropanol and acetaldehyde and acetone from isopropanol oxidation
CO2
Isopropil alcohol
Methanol was emitted
by leaves
formaldehyde
and
formic acid was not
detected.
After
ethanol
feeding
acetaldehyde was the most
abundant
organic
compound emitted trough
stomata.
PTR-MS
showed
that
Isopropyl
alcohol
was
adsorbed by rocket plants,
translocated and re-emitted
by rocket leaves.
Isopropyl alcohol was also
metabolized in acetaldehyde
and in acetone reemitted in
the atmosphere.
Conclusions
• An array of measuring techniques should be used to track the fate of each organic
compound when it enters in contact with a plant via the nutrient solution delivery system.
• Even similar molecules can have a very different fate when in contact with plants.
• Plant metabolism can destroy the pollutant or even produce other equally or even more
dangerous molecules and even release them in the atmosphere.
• There is no general rule to be applied. Specific interaction plant/pollutant should be tested
with reference to the growing conditions.
Perspectives
An accurate definition of organic pollutants that can enter in a plant growing system in
Bioregenerative Life Support system is necessary to ensure :
plant growing and productive performances.
food quality and security.
the potential treat of release in the system of the same or derived pollutants.