astrobiology.gsfc.nasa/dworkin/

Analysis of natural and synthetic extraterrestrial material Jason Dworkin Meteorites Returned Samples Ice Simulations Grain Simulations http://astrobiology.gsfc.nasa.gov/dworkin/

Observations & Simulations Lab Analysis Simulations

Spectroscopy (e.g. IR) Is Used for Remote Observations Scott Sandford, NASA ARC

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

Chromatographic Tools Liquid Chromatography Mass Spectrometry LC/MS Gas Chromatography Mass Spectrometry GC/MS HPLC & Detectors MS MS GC nanoLC LIF Detector

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Dworkin Co-Authored AbSciCon Presentations • Investigation of isovaline enantiomeric excesses in CM meteorites using liquid chromatography time of flight mass spectrometry. Glavin & Dworkin • Reassessing the organic content of antarctic ice and meteorites. Botta et al. • Icy prebiotic chemistry of nitriles and other molecules. Hudson, Moore & Dworkin • Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila, Bernstein & Dworkin • A possible pathway for organic synthesis and implications for protostellar systems. Johnson et al. • Prebiotic phosphorylation through phosphorus radicals. Pasek, Dworkin & Lauretta • MALDI-MS analysis of hetero-trimer fractions formed by montmorillonite catalysis in the reaction of binary monomer mixtures. Hazen (Ertem) et al. • 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

gly gABA AIB bala ala bABA Synergies: Amino Acids Collaborations 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

Amino Acid Protocol Sample Water extraction (100ºC 24 h) 50% Acid hydrolysis (6 M HCl 150ºC 3 h) HPLC with UV fluorescence+ ToF-MS detection Desalting of soils or meteorites (AG50W-X8 resin) 50% Derivatization (OPA/NAC primary amines)

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. hn 10 K H2O+CH3OH+HCN+NH3 Blank Fluorescence Gly Standard Fluorescence b Ala g ABA Ala D L b ABA D L Sample Fluorescence

Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila et al. hn 10 K H2O+CH3OH+HCN+NH3 Negative electrospray OPA/NAC-Serine (-H+) 1 13C 2 13C Positive electrospray OPA/NAC-Serine (+H+) 1 13C 2 13C m/z Mass calibrate to internal or external standard

Studies of amino acid formation by photolysis of interstellar ice analogs. Elsila et al. Alanine & b Alanine ESI+ Glycine ESI+ Serine ESI+ Fluorescence

Standards C6 C5 C4 C3 C2 C3OH C3COOH C2COOH

Murchison Meteorite, CM2 Danny Glavin’s Talk

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

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. 1/2 L sampling bottle containing air from near the landing site vents analyzed at JSC. • The Sample Canister Filter has yet to be analyzed for any trapped gases.

Analytical techniques likely to be used for analysis of returned samples: Analytical techniques 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) None will detect specific biomolecules! Interplanetary Dust Particle (IDP) Comet Wild 2

Can LC/MS techniques used in biotechnology be applied? 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 (b 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 Interplanetary Dust Particle (IDP) Comet Wild 2

Stardust or IDP (10-20 µg) ToF-MS Water extraction (100ºC 24 h) 50% LIF Acid hydrolysis (6 M HCl 150ºC 3 h) 50% Derivatization (OPA/NAC primary amines) nLC nLC with LIF + nanospray ToF-MS detection

Separation Column: 70µm x 100 mm 1.7µm resin

Fluorescence Detection Diode pulsed laser 70µm flow cell PMT detector

Ionization Reference Sprayer Baffle MS Inlet Sample 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 HPLC 4.6 mm, 5 µm resin 2 mm, 3 µm resin 1 mm, 3 µm resin nLC 300 µm, 3 µm resin 100 µm, 3 µm resin 100 µm, 1.7 µm resin  70 µm, 1.7 µm resin 50 µm, 3 µm resin & pre-column 70 µm, 1.7 µm resin Goal Phenylhexyl XTerra C18 BEH C18

CI Meteorites: A Cometary Origin? CI CM Orgueil (France, 1864) AIB b-alanine glycine • 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 Ehrenfreund et al. PNAS 98 (2001) 2138-2141

Ala • Ala AIB • ABA • ABA g ABA

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