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Determination of (alpha)-dialkylamino acids and their Enantiomers in Geological Samples by High-Performance Liquid Chromatography after Dervatization with a Chiral Adduct of (omicron)-Phthaldialdehyde

Derivatization with (omicron)-phthaldialdehyde (OPA) and the chiral thiol N-acetyl-L-cysteine (NAC) is a convenient and sensitive technique for the HPLC detection and resolution of protein amino acid enantiomers. The kinetics of the reaction of OPA-NAC with (alpha)-dialkylamino acids was investigated. The fluorescence yield of (alpha)-dialkylamino acids was only about 10% of that of protein amino acids when the derivatization was carried out at room temperature for 1-2 min, which is the procedure generally used for protein amino acid analyses. The fluorescence yield of (alpha)-dialkylamino acids can be enhanced by up to ten-fold when the derivatization reaction time is increased to 15 min at room temperature. The OPA-NAC technique was optimized for the detection and enantiomeric resolution of a-dialkylamino acids in geological samples which contain a large excess of protein amino acids. The estimated detection limit for a-dialkylamino acids is 1-2 pmol, comparable to that for protein amino acids.

Zhoa, Meixun

A Reexamination of Amino Acids in Lunar Soils: Implications for the Survival of Exogenous Organic Material During Impact Delivery

Using a sensitive high performance liquid chromatography technique, we have analyzed both the hot water extract and the acid hydrolyzed hot water extract of lunar soil collected during the Apollo 17 mission. Both free amino acids and those derived from acid labile precursors are present at a level of roughly 15 ppb. Based on the D/L amino acid ratios, the free alanine and aspartic acid observed in the hot water extract can be entirely attributed to terrestrial biogenic contamination. However, in the acid labile fraction, precursors which yield amino acids are apparently present in the lunar soil. The amino acid distribution suggests that the precursor is probably solar wind implanted HCN. We have evaluated our results with regard to the meteoritic input of intact organic compounds to the moon based on an upper limit of less than or equal to 0.3 ppb for alpha-aminoisobutyric acid, a non-protein amino acid which does not generally occur in terrestrial organisms and which is not a major amino acid produced from HCN, but which is a predominant amino acid in many carbonaceous chondrites. We find that the survival of exogenous organic compounds during lunar impact is less than or equal to 0.8%. This result represents an example of minimum organic impact survivability. This is an important first step toward a better understanding of similar processes on Earth and on Mars, and their possible contribution to the budget of prebiotic organic compounds on the primitive Earth.

Brinton, Karen L. F.

Investigation of the Prebiotic Synthesis of Amino Acids and RNA Bases from CO2 Using FeS/H2S As a Reducing Agent

An autotrophic theory of the origin of metabolism and life has been proposed in which carbon dioxide is reduced by ferrous sulfide and hydrogen sulfide by means of a reversed citric acid cycle, leading to the production of amino acids. Similar processes have been proposed for purine synthesis. Ferrous sulfide is a strong reducing agent in the presence of hydrogen sulfide and can produce hydrogen as well as reduce alkenes, alkynes, and thiols to saturated hydrocarbons and reduce ketones to thiols. However, the reduction of carbon dioxide has not been demonstrated. We show here that no amino acids, purities, or pyrimidines are produced from carbon dioxide with the ferrous sulfide and hydrogen sulfide system. Furthermore, this system does not produce amino acids from carboxylic acids by reductive amination and carboxylation. Thus, the proposed autotrophic theory, using carbon dioxide, ferrous sulfide, and hydrogen sulfide, lacks the robustness needed to be a geological process and is, therefore, unlikely to have played a role In the origin of metabolism or the origin of life.

Keefe, Anthony D.

Detecting Amino Acids on Mars

Understanding the events that led to the origin of life on Earth is complicated by the lack of geological evidence from the period around four billion years (4 Gyr) ago when the transition from prebiotic chemistry to biochemistry is believed to be occurred. Although erosion and plate tectonics have since erased the terrestrial geological record from the time of the origin of life, there is possibility that information about this period of Earth history may still be preserved on Mars. A major goal of the NASA Space Exploration Program is to search for evidence of abiotic chemistry and extinct or extant life on Mars. During the next decade, spacecraft will orbit Mars, land on the surface, and return with surface samples for analysis. The question is what compounds should we search for, either directly on the planet or in samples returned to Earth, that will answer unambiguously whether abiotic and/or biotic organic molecules are present.

Bada, Jeffrey L.

A Search for Extraterrestrial Amino Acids in Polar Ice: A Progress Report

Fifteen polar ice samples-fourteen from Greenland and one from Antarctica-have been analyzed for the extraterrestrial amino acid alpha-aminoisobutyric acid (AIB) in an effort to estimate the flux of interplanetary organic material to the Earth's surface. Only one sample (Greenland GISP II, 4270-4440 years old) contains detectable amounts of AIB, apparently the signature of a transient delivery event. The maximum oceanic concentration of AIB from such an event would be less than 10(exp-9) M.

Bada, Jeffrey L.

The Chemistry of Early Self-Replicating Systems

The NASA Specialized Center of Research and Training (NSCORT) in Exobiology is a consortium of scientists at the University of California at San Diego (UCSD), The Salk Institute for Biological Studies (Salk) and The Scripps Research Institute (TSRI). All three institutions are located in close geographical proximity in La Jolla, California. The NSCORT/Exobiology is administered through the Scripps Institution of Oceanography. Since its inception in January 1992, the NSCORT in Exobiology has made major contributions with respect to the question of how life began on Earth. The Principal Investigators (PIs) and their associated Fellows have published numerous articles in peer reviewed journals on topics relevant to Exobiology. They have presented papers and sponsored symposia at several meetings of national and international scientific societies. A total of 30 undergraduate, 12 graduate and 15 postdoctoral Fellows have been supported by the NSCORT. The Fellows have met on their own at least once a month to discuss Exobiology topics and research progress. The NSCORT has arranged seminars and evening discussion meetings, and offered an undergraduate class on "Biochemical Evolution" as well as graduate courses dealing with topics in Exobiology. A visiting scientist program has allowed 11 scientists from the U.S. and 4 foreign countries to conduct cooperative research with the various PIs. An active outreach program has been initiated, which includes an Exobiology high school level teaching module and curriculum guide, and an elementary school level booklet about basic atomic structure and formation of the universe, Sun and Earth. A World Wide Web Homepage (http://www-chem.ucsd.edu/-nscort/ NSCORT.html) has been developed, which describes the NSCORT activities, research programs and Fellowship opportunities. The various activities of the NSCORT in Exobiology have received wide-spread coverage in both the scientific and public media. The major function of the NSCORT is the training of young scientists in the field of Exo- biology. Thus, the bulk of the $1,000,000 annual budget is used to support the research and training of undergraduate, graduate and post-doctoral Fellows who are selected on a competitive basis. About five Fellows at each level are supported each year. Our goal is to train scientists whose major research interest is Exobiology, but whose mastery in the classical fields of chemistry, biology and earth science is so strong that they outstanding candidates for either graduate school or academic tenure-track positions in departments at leading national and international Universities. Applicants for these Fellowships are solicited by advertisements in journals such as Science and Nature and in organizational newsletters such as the one published by the International Society for the Study of the Origin of Life (ISSOL), by contacting academic and NASA colleagues working in Exobiology or related fields and by recruiting students who have already been admitted into the various academic programs with which the PIs are affiliated.

Bada, Jeffrey L.

Investigation of the Prebiotic Synthesis of Amino Acids and RNA Bases from CO2 using FeS/H2S as a Reducing Agent

An autotrophic theory of the origin of metabolism and life has been proposed in which carbon dioxide is reduced by ferrous sulfide and hydrogen sulfide by means of a reversed citric acid cycle, leading to the production of amino acids. Similar processes have been proposed for purine synthesis. Ferrous sulfide is a strong reducing agent in the presence of hydrogen sulfide and can produce hydrogen as well as reduce alkenes, alkynes, and thiols to saturated hydrocarbons and reduce ketones to thiols. However, the reduction of carbon dioxide has not been demonstrated. We show here that no amino acids, purines, or pyrimidines are produced from carbon dioxide with the ferrous sulfide and hydrogen sulfide system. Furthermore, this system does not produce amino acids from carboxylic acids by reductive amination and carboxylation. Thus, the proposed autotrophic theory, using carbon dioxide, ferrous sulfide, and hydrogen sulfide, lacks the robustness needed to be a geological process and is, therefore, unlikely to have played a role in the origin of metabolism or the origin of life.

Keefe, Anthony D.

Mars Sample Handling and Requirements Panel (MSHARP)

In anticipation of the return of samples from Mars toward the end of the first decade of the next century, NASA's Office of Space Sciences chartered a panel to examine how Mars samples should be handled. The panel was to make recommendations in three areas: (1) sample collection and transport back to Earth; (2) certification of the samples as nonhazardous; and (3) sample receiving, curation, and distribution. This report summarizes the findings of that panel. The samples should be treated as hazardous until proven otherwise. They are to be sealed within a canister on Mars, and the canister is not to be opened until within a Biosafety Hazard Level 4 (BSL-4) containment facility here on Earth. This facility must also meet or exceed the cleanliness requirements of the Johnson Space Center (JSC) facility for curation of extraterrestrial materials. A containment facility meeting both these requirements does not yet exist. Hazard assessment and life detection experiments are to be done at the containment facility, while geochemical characterization is being performed on a sterilized subset of the samples released to the science community. When and if the samples are proven harmless, they are to be transferred to a curation facility, such as that at JSC.

Carr, Michael H.

Fullerenes in Allende Meteorite

The detection of fullerenes in deposits from meteor impacts has led to renewed interest in the possibility that fullerenes are present in meteorites. Although fullerenes have not previously been detected in the Murchison and Allende meteorites, the Allende meteorite is known to contain several well-ordered graphite particles which are remarkably similar in size and appearance to the fullerene-related structures carbon onions and nanotubes. We report that fullerenes are in fact present in trace amounts in the Allende meteorite. In addition to fullerenes, we detected many polycyclic aromatic hydrocarbons (PAHs) in the Allende meteorite, consistent with previous reports. In particular, we detected benzofluoranthene and corannulene (C20H10), five-membered ring structures which have been proposed as precursors to the formation of fullerene synthesis, perhaps within circumstellar envelopes or other sites in the interstellar medium.

Becker, L.

MOD: An Organic Detector for the Future Exploration of Mars

The Mars Organic Detector (MOD) is designed to assess whether organic compounds, possibly associated with life, are present in Martian rock and soil samples. MOD has a detection limit that is at least two orders of magnitude more sensitive than the Viking GCMS. MOD is focused on detecting amino acids, amines and PAH (polycyclic aromatic hydrocarbons). Amino acids play an essential role in biochemistry on Earth and PAH are widespread throughout the universe and can provide an indication of the delivery of meteoritic organic material to Mars. The advantage of MOD is the absence of wet chemistry and its simple and robust design. The sample will be extracted from the mineral matrix (0.1 - 1 g of rock-powder) using sublimation and analyzed with a fluorescence detector. The isolation method is based on the fact that amino acids and PAH are volatile at temperatures greater than 150C. The fluorescence detection scheme is based on UV excitation with LED's, optical filters, PrN diode photon detector and a sample calibration reservoir. Fluorescamine is used as a fluorescing reagent for amino acids and amines, while PAH are naturally fluorescent. There is no sample preparation required and the turnaround time for a single analysis is on the order of minutes.

Kminek, G.

Extraterrestrial Helium (He@C60) Trapped in Fullerenes in the Sudbury Impact Structure

Fullerenes (C60 and C70) have recently been identified in a shock-produced breccia (Onaping Formation) associated with the 1.85-Ga Sudbury Impact Crater. The presence of parts-per-million levels of fullerenes in this impact structure raises interesting questions about the processes that led to the formation of fullerenes and the potential for delivery of intact organic material to the Earth by a large bolide (e.g., asteroid or comet). Two possible scenarios for the presence of fullerenes in the Sudbury impact deposits are that (1) fullerenes are synthesized within the impact plume from the C contained in the bolide; or (2) fullerenes are already present in the bolide and survived the impact event. The correlation of C and trapped noble gas atoms in meteorites is well established. Primitive meteorites contain several trapped noble gas components that have anomalous isotopic compositions, some of which may have a presolar origin. Several C-bearing phases, including SiC, graphite, and diamond, have been recognized as carriers of trapped noble gases. It has also been suggested that fullerenes (C60 and C70) might be a carrier of noble gas components in carbonaceous chondrites. Recently, fullerenes have been detected in separate samples in the Allende meteorite. Carbon-60 is large enough to enclose the noble gases He, Ne, Ar, Kr, and Xe, but it is too small to contain diatomic gases such as N2 or triatomic gases such as CO2. Recent experimental work has demonstrated that noble gases of a specific isotopic composition can be introduced into synthetic fullerenes at high temperatures and pressures; these encapsulated gases can then be released by the breaking of one or more C bonds during step-heating under vacuum. These thermal-release patterns for He encapsulated within the C60 molecule (He@C60) are similar to the patterns for acid residues of carbonaceous chondrites, suggesting that fullerenes could be an additional carrier of trapped noble gases in acid residues of meteorites. Analysis and Results: In order to characterize the noble gas compositions of the Sudbury fullerenes, we undertook a systematic study of acid-resistant residues throughout the C-rich layer (Black member) of the Onaping Formation. Samples were demineralized and extracted using standard techniques. The Onaping extracts were analyzed using several techniques, including UV-Vis adsorption, electro spray mass spectrometry, and laser desorption (linear and reflectron) time-of-flight (TOF) mass spectrometry (LDMS). The Sudbury fullerenes were then separated and purified using HPLC coupled with a photo diode array detector. The HPLC extracts containing the purified fullerenes were loaded into a metal tube furnace within a glove box under a N atmosphere in preparation for noble gas analyses. The 3-He and 4-He content of the fullerene extracts was measured using previously reported standard techniques . Discussion: Fullerenes (C60 and C70) in the Sudbury Impact Structure have been found to contain trapped He with a 3-He/4-He ratio greater than 5 x 10(exp -4). The 3-He/4-He ratio exceeds the accepted solar value by more than 30% and is more than 10x higher than the maximum reported mantle value. Terrestrial nuclear reactions or cosmic-my bombardment are not sufficient to generate such a high ratio. The 3-He/4-He ratios in the Sudbury fullerenes are similar to those determined for interplanetary dust particles. The greater-than-solar ratios of 3-He/4-He in the Sudbury fullerenes may indicate a presolar origin, although alternative mechanisms occurring in the ISM to explain these high ratios (e.g., spallation reactions, selective He implantation, etc.) cannot be entirely ruled out. We are currently attempting to isolate enough fullerene material to measure anomalous Ne (or Kr or Xe) contained within the C60 (e.g., the "pure" 22-Ne component) and thus determine whether the Sudbury fullerenes are indeed presolar in origin.

Becker, L.

Integrated Microchemical Analysis System Using DS2 Penetrator Technology for the Enantiomeric Detection of Amino Acids

Any strategy for investigating whether abiotic and/or biotic organic molecules are present on Mars and the search for biosignatures should focus on compounds which are readily synthesized under plausible prebiotic conditions, play an essential role in biochemistry as we know it and have properties such as chirality (handedness) which can be used to distinguish between abiotic vs. biotic origins (1). Amino acids are one of the few compound classes that fulfill all these requirements. They are synthesized in high yields in prebiotic simulation experiments, are one of the more abundant types of organic compounds present in carbonaceous meteorites and only the L-enantiomers are used in the proteins and enzymes in life on Earth.

Frank J Grunthaner

A Cometary Origin of the Amino Acids in the Orgueil Meteorite?

A reexamination of a piece of the Orgueil meteorite revealed that its amino acid composition is strikingly different to two other carbonaceous chondrites, suggesting different parent bodies. A cometary origin for Orgueil would be one possibility.

Botta, O.

MOD: An In-Situ Organic Detector for the MSR 2003 Mission

Looking for organic compounds that are essential for biochemistry or indicative of extraterrestrial organic influx is the primary goal of MOD (Mars Organic Detector). MOD can also quantify adsorbed and chemisorbed water and evolved carbon dioxide.

Kminek, G.

Amino Acid Chemistry as a Link Between Small Solar System Bodies and Carbonaceous Chondrites

Establishing chemical links between meteorites and small solar system bodies, such as comets and asteroids, provides a tool for investigating the processes that occurred during the formation of the solar system. Carbonaceous meteorites are of particular interest, since they may have seeded the early Earth with a variety of prebiotic organic compounds including amino acids, purines and pyrimidines, which are thought to be necessary for the origin of life. Here we report the results of high-performance liquid chromatography (HPLC) based amino acid analyses of the acid-hydrolyzed hot water extracts from pristine interior pieces of the CI carbonaceous chondrites Orgueil and Ivuna and the CM meteorites Murchison and Murray. We found that the CI meteorites Orgueil and Ivuna contained high abundances of beta-alanine and glycine, while only traces of other amino acids like alanine, alpha-amino-n-butryic acid (ABA) and alpha-aminoisobutyric acid (AIB) were detected in these meteorites. Carbon isotopic measurements of beta-alanine and glycine in Orgueil by gas chromatography combustion-isotope ratio mass spectrometry clearly indicate an extraterrestrial origin of these amino acids. The amino acid composition of Orgueil and Ivuna was strikingly different from the CM chondrites Murchison and Murray. The most notable difference was the high relative abundance of B-alanine in Orgueil and Ivuna compared to Murchison and Murray. Furthermore, AIB, which is one of the most abundant amino acids found in Murchison and Murray, was present in only trace amounts in Orgueil and Ivuna. Our amino acid data strongly suggest that the CI meteorites Orgueil and Ivuna came from a different type of parent body than the CM meteorites Murchison and Murray, possibly from an extinct comet. It is generally thought that carbonaceous meteorites are fragments of larger asteroidal bodies delivered via near Earth objects (NEO). Orbital and dynamic studies suggest that both fragments of main belt asteroids and comets replenish the NEO population, therefore extinct comets may contribute up to half of all NEO's. A comparison of an amino acid analysis of a returned NEO sample to CI and CM carbonaceous chondrites would help establish a link between small solar system bodies and meteorites. Based on our amino acid measurements of CI and CM chondrites, amino acid chemistry can be included as an additional set of criteria to constrain the nature of meteorite parent bodies.

Glavin, Daniel P.

The Next Generation MOD: A Microchip Amino Acid Analyzer for Detecting Extraterrestrial Life

The MOD (Mars Organic Detector) instrument which has selected for the definition phase of the BEDS package on the 2005 Mars Explorer Program spacecraft is designed to simply detect the presence of amino acids in Martian surface samples at a sensitivity of a few parts per billion (ppb). An additional important aspect of amino acid analyses of Martian samples is identifying and quantifying which compounds are present, and also distinguishing those produced abiotically from those synthesized by either extinct or extant life. Amino acid homochirality provides an unambiguous way of distinguishing between abiotic vs. biotic origins. Proteins made up of mixed D- and L-amino acids would not likely have been efficient catalysts in early organisms because they could not fold into bioactive configurations such as the a-helix. However, enzymes made up of all D-amino acids function just as well as those made up of only L-amino acids, but the two enzymes use the opposite stereoisomeric substrates. There are no biochemical reasons why L-amino acids would be favored over Damino acids. On Earth, the use of only L-amino acids in proteins by life is probably simply a matter of chance. We assume that if proteins and enzymes were a component of extinct or extant life on Mars, then amino acid homochirality would have been a requirement. However, the possibility that Martian life was (or is) based on D-amino acids would be equal to that based on L-amino acids. The detection of a nonracemic mixture of amino acids in a Martian sample would be strong evidence for the presence of an extinct or extant biota on Mars. The finding of an excess of D-amino acids would provide irrefutable evidence of unique Martian life that could not have been derived from seeding the planet with terrestrial life (or the seeding of the Earth with Martian life). In contrast, the presence of racemic amino acids, along with non-protein amino acids such as alpha-aminoisobutyric acid and isovaline, would be indicative of an abiotic origin, although we have to consider the possibility that the racemic amino acids were generated from the racemization of biotically produced amino acids.

Mathies, R. A.

MOD: An Instrument for the 2005 Mars Explorer Program HEDS Payload

The Mars Organic Detector (MOD) was recently selected for the definition phase of the HEDS '05 (originally scheduled for '03) lander instrument package for fundamental biology and in situ resource utilization. MOD is designed to detect organic compounds in rock and soil samples directly on the surface of Mars in order to assess the biological potential of the planet. In addition, a MOD Tunable Diode Laser Spectrometer (TDLS) will provide information on desorption and decomposition temperatures, as well as the release rates and quantities of water and carbon dioxide that can be liberated from regolith samples, thereby providing the parameters needed for the design of systems for the future large-scale in situ extraction of valuable consumable resources. A MOD TDLS will also measure the atmospheric water and carbon dioxide content, as well as the atmospheric carbon dioxide isotopic composition, in order to determine whether there is an isotopic offset between atmospheric and surface carbon.

Bada, J. L.

Integrated Micro-Chip Amino Acid Chirality Detector for MOD

Integration of a micro-chip capillary electrophoresis analyzer with a sublimation-based extraction technique, as used in the Mars Organic Detector (MOD), for the in-situ detection of amino acids and their enantiomers on solar system bodies. Additional information is contained in the original extended abstract.

Glavin, D. P.