http://astrobiology.gsfc.nasa.gov/dworkin/

Meteorites

Ice Simulations

Grain Simulations

Returned Samples

Analysis of natural and synthetic extraterrestrial material Jason Dworkin

Observations & Simulations

Lab Analysis

Simulations

Scott Sandford, NASA ARC

Spectroscopy (e.g. IR) Is Used for Remote Observations

Russian botanist Mikhail Tsvet invented chromatography in 1901 during his research on chlorophyll.

Chromatographic Tools

HPLC & Detectors

nanoLC LIF Detector

MS

Liquid ChromatographyMass Spectrometry

LC/MS

Gas ChromatographyMass Spectrometry

GC/MS

MS GC

(NAI)

(NAI)

(NAI)

(NAI)

(NAI)

Dworkin Co-Authored AbSciCon Presentations1. Investigation of isovaline enantiomeric excesses in CM meteorites

using liquid chromatography time of flight mass spectrometry. Glavin & Dworkin

2. Reassessing the organic content of antarctic ice and meteorites. Botta et al.

3. Icy prebiotic chemistry of nitriles and other molecules. Hudson, Moore & Dworkin

4. Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila, Bernstein & Dworkin

5. A possible pathway for organic synthesis and implications for protostellar systems. Johnson et al.

6. Prebiotic phosphorylation through phosphorus radicals. Pasek, Dworkin & Lauretta

7. MALDI-MS analysis of hetero-trimer fractions formed by montmorillonite catalysis in the reaction of binary monomer mixtures. Hazen (Ertem) et al.

8. The astrobiology in secondary classroom (ASC) project. Walter, et al.

• STARDUST amino acid analysis • Hydrazine thruster contamination

• Amino acids in CM & CR meteorites • SAM GC/MS

• Nucleobases in CM meteorites • Asteroid sample return mission

1. Investigation of isovaline enantiomeric excesses in CM meteorites using liquid chromatography time of flight mass spectrometry. Glavin & Dworkin

2. Reassessing the organic content of antarctic ice and meteorites. Botta et al.

3. Icy prebiotic chemistry of nitriles and other molecules. Hudson, Moore & Dworkin

4. Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila, Bernstein & Dworkin

5. A possible pathway for organic synthesis and implications for protostellar systems. Johnson et al.

6. Prebiotic phosphorylation through phosphorus radicals. Pasek, Dworkin & Lauretta

7. MALDI-MS analysis of hetero-trimer fractions formed by montmorillonite catalysis in the reaction of binary monomer mixtures. Hazen (Ertem) et al.

8. The astrobiology in secondary classroom (ASC) project. Walter, et al.

• STARDUST amino acid analysis • Hydrazine thruster contamination

• Amino acids in CM & CR meteorites • SAM GC/MS

• Nucleobases in CM meteorites • Asteroid sample return mission

Dworkin Co-Authored AbSciCon Presentations

Synergies: Amino AcidsCollaborations with Glavin and Botta have focused research on amino acids and

meteorites.

While I am fond of saying, “There is more to life than amino acids,” these compounds are relevant, popular, and (with our method of analysis) easy to generate extraordinary data.

1/2 of the AbSciCon abstracts from this lab are based on amino acid data from meteorites or lab simulations.

Lab facilities have been used to help characterize SAM derivitization agent

glyala

ABAABAAIB

ala

Amino Acid Protocol

Sample

Water extraction (100ºC 24 h)

Acid hydrolysis (6 M HCl 150ºC 3 h)

Desalting of soils or meteorites

(AG50W-X8 resin)

Derivatization (OPA/NAC primary amines)

HPLC with UV fluorescence+ ToF-MS detection

50%

50%

chirality(non-super imposable mirror images)

With few exceptions, life is homochiral and abiotic chemistry is racemic

Using a chiral fluorescent label forms diastereomers with chiral amino acids (L,L and D,L) and allows for separation.

Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila et al.

Standard Fluorescence

Sample Fluorescence

Blank Fluorescence

Gly

ABAD L

ABA Ala

AlaD L

H2O+CH3OH+HCN+NH3

h10 K

Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila et al.

H2O+CH3OH+HCN+NH3

h10 K

m/z

Positive electrospray

Negative electrospray

OPA/NAC-Serine (-H+)

OPA/NAC-Serine (+H+)

1 13C2 13C

1 13C

2 13C

Mass calibrate to internal or external standard

Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila et al.

Serine ESI+

Fluorescence

Glycine ESI+

Alanine & Alanine ESI+

C2

C3

C4

C5

C6

C3OH

C3COOH

C2COOH

Standards

Danny Glavin’s Talk

Murchison Meteorite, CM2

Member of Organics Preliminary Examination Team

• Analyze landing site mud for amine contamination• Arrange analysis of landing site and clean room air• Analyze heat shield and filters for amines• Analyze flight aerogel for amine contamination• Analyze comet-exposed aerogel for amines

STARDUST - Evaluation of Sources of 1° Amine Contamination (R-NH2)

Several flight quality aerogels and UTTR soil and standing water collected 2004 from near the Genesis recovery site were analyzed for amino acids. The results are compared to the Murchison (CM) and Orgueil (CI) meteorites.

abundance (ppm) Aerogel Soil Water Murchison Orgueil

Amines 7.0 12.0 2.0 14.6 4.2

Most Abundant

(ppm)

Nylon*

5.4

L-glutamic acid

2.4

-alanine

0.3

AIB**

3.2

-alanine

2.1

AIB** <0.01 <0.006 <0.004 3.2 0.04

-Alanine 0.03 0.2 0.3 1.4 2.1

*-amino-n-caproic acid, from Nylon-6 degradation **-amino-isobutyric acid

Aerogel contamination should not be a problem for the STARDUST analyses. While, it is not anticipated that the samples were exposed to UTTR soil or water, some compounds (e.g. AIB) could still be determined.

Analysis of air samples from the landing site and UTTR clean room

•Air samples were taken at the landing site near the heat shield and at the vents and in the UTTR clean room near the heat shield and the interior of the SRC.

•Analyzed by GC and GC/MS at the NASA JSC Toxicology Laboratory.

•Trace levels of volatile organic compounds were found at the heat shield and vents at the landing site.

•Isopropanol and 1,1,1,2-tetrafluoroethane were found in the ppm range as well as trace levels of other volatile organic compounds at the heat shield and the interior of the SRC, after transfer to the Utah clean room.

•The Sample Canister Filter has yet to be analyzed for any trapped gases.

1/2 L sampling bottle containing air from near the landing site vents analyzed at JSC.

Analytical techniques likely to be used for analysis of returned samples:

NanoSIMS Ion Microprobe): elemental and isotopic composition map

Inductively Coupled Plasma Mass Spectrometry (ICP-MS): elemental and isotopic composition

X-ray Absorption Near Edge Structure (XANES) Spectroscopy: general bonding of organics

Micro Laser Desorption Ionization Mass Spectrometry (µL2MS): polycyclic aromatic hydrocarbons (PAHs)

Comet Wild 2 Interplanetary Dust Particle (IDP)

None will detect specific biomolecules!

Analytical techniques to be used for analysis of returned samples

Comet Wild 2 Interplanetary Dust Particle (IDP)

Mass of single 10 µm grain: ~ 1 x 10-9 g

Concentration of AIB:

Murchison meteorite 3 ppm (µg/g)

Antarctic µmeteorite 0.2 ppm

Orgueil meteorite 0.04 ppm

Orgueil meteorite ( Ala) 2 ppm

Molecules AIB/grain (x 10-18 moles): 80 to 0.5

Detection limit (x 10-18 moles): ~10

Number 10 µm grains required: 1/4 to 20

Number 20 µm grains required: 1/32 to 2

Can LC/MS techniques used in biotechnology be applied?

Stardust or IDP

(10-20 µg)

Water extraction (100ºC 24 h)

Acid hydrolysis (6 M HCl 150ºC 3 h)

Derivatization (OPA/NAC primary amines)

nLC with LIF

+ nanospray ToF-MS detection

50%

50%

nLC

ToF-MS

LIF

Separation

Column:70µm x 100 mm1.7µm resin

Fluorescence Detection

Diode pulsed laser70µm flow cellPMT detector

Ionization

Reference SprayerBaffleMS InletSample Sprayer (20µm)

Detection of 10 amol of AIB by MS

To do:• Reduce contamination (glass pipettes, pyrolyzed salts, clean room?)• Adapt HPLC separation technique to nLC

Adapt HPLC separation technique to nLC

4.6 mm, 5 µm resin

2 mm, 3 µm resin

1 mm, 3 µm resin

300 µm, 3 µm resin

100 µm, 3 µm resin

100 µm, 1.7 µm resin

70 µm, 1.7 µm resin

HPLC

nLC

Phenylhexyl XTerra C18 BEH C18

70 µm, 1.7 µm resin50 µm, 3 µm resin & pre-column Goal

CI CM

• CIs: fragments of extinct cometary nuclei (Lodders and Osborne, 1999)

• Amino acid composition in CIs distinct from CMs

• Amino acids consistent with volatiles detected in comets Hyakutake and Hale-Bopp (Crovisier and Bockelée-Morvan, 1999)

• Direct analysis of comet or asteroid will help constrain the nature of meteorite parent bodies

CI Meteorites: A Cometary Origin?

Orgueil (France, 1864)

glycine

-alanineAIB

Ehrenfreund et al. PNAS 98 (2001) 2138-2141

Ala Ala

AIB ABA ABA

ABA

QuickTime™ and aTIFF (Uncompressed) decompressor

are needed to see this picture.

Ala Ala

AIB ABA ABA

ABA

QuickTime™ and aTIFF (Uncompressed) decompressor

are needed to see this picture.