AJAC  Vol.6 No.4 , March 2015
A Method for Detection of Trace Concentrations of Underivatized Amino Acid in Hydrothermal Fluids by Ion-Pairing Reversed-Phase UPLC-ESI-QTOF-MS
Abstract: Investigation of amino acids in hydrothermal systems is of prime importance for the understanding of geochemistry and microbiology of hydrothermal vents and plumes, for carbon and metals global cycles, for metabolism of some hydrothermal microorganisms and for the origin of life issue. Extensive theoretical and experimental work on amino acids behaviour in hydrothermal fluids has been done, conversely only few data exist on natural samples. Because each hydrothermal vent is unique, the more data we collect the better we will be able to address each of these questions. Usually amino acids in hydrothermal fluids have been measured by HPLC-FLD. The chromatographic separation was at least 26 min and up to 135 min and the required derivatization step may be time consuming, may use harmful chemicals and may be source of contamination. Alternatively, we describe here a method combining quickness (4.5 min), high resolution (10,000), very low LOD (sub-ppb) and without derivatization. Characterisation and separation of 10 relevant proteinogenic underivatized amino acids was achieved by ion-pairing reversed-phase Ultra-high Performance Liquid Chromatography-Electrospray Ionisation-Quadrupole Time of Flight-Mass Spectrometry (UPLC-ESI-QTOF-MS). Excellent linearity in the response was obtained for all amino acids with correlation coefficients > 0.9921. This method was successfully applied to natural hydrothermal fluid samples from ultramafic-hosted vents of the Mid-Atlantic Ridge region. Results are consistent with the only 2 other studies published on ultramafic-hosted vents and complete the few available data.
Cite this paper: Konn, C. , Magnér, J. , Charlou, J. , Holm, N. and Alsberg, T. (2015) A Method for Detection of Trace Concentrations of Underivatized Amino Acid in Hydrothermal Fluids by Ion-Pairing Reversed-Phase UPLC-ESI-QTOF-MS. American Journal of Analytical Chemistry, 6, 313-324. doi: 10.4236/ajac.2015.64030.

[1]   Corliss, J.B., Baross, J.A. and Hoffman, S.E. (1981) An Hypothesis Concerning the Relationship between Submarine Hot Springs and the Origin of Life on Earth. Proceedings 26th International Geological Congress, Oceanologica Acta, No SP, 59-69.

[2]   Holm, N.G. (1992) Chapter 1 Why Are Hydrothermal Systems Proposed as Plausible Environments for the Origin of Life? Origins of Life and Evolution of Biospheres, 22, 5.

[3]   Shock, E.L. (1992) Chapter 5 Chemical Environments of Submarine Hydrothermal Systems. Origins of Life and Evolution of Biospheres, 22, 67-107.

[4]   Ferris, J.P. (1992) Chapter 6 Chemical Markers of Prebiotic Chemistry in Hydrothermal Systems. Origins of Life and Evolution of Biospheres, 22, 109-134.

[5]   Macleod, G., McKeown, C., Hall, A.J. and Russell, M.J. (1994) Hydrothermal and Oceanic pH Conditions of Possible Relevance to the Origin of Life. Origins of Life and Evolution of Biospheres, 24, 19-41.

[6]   Shock, E. and Canovas, P. (2010) The Potential for Abiotic Organic Synthesis and Biosynthesis at Seafloor Hydrothermal Systems, Blackwell Publishing Ltd., 161-192.

[7]   Shock, E.L. (1990) Geochemical Constraints on the Origin of Organic Compounds in Hydrothermal Systems. Origins of Life and Evolution of Biospheres, 20, 331-367.

[8]   Aubrey, A., Cleaves, H. and Bada, J. (2009) The Role of Submarine Hydrothermal Systems in the Synthesis of Amino Acids. Origins of Life and Evolution of Biospheres, 39, 91-108.

[9]   McCollom, T.M. (2013) Laboratory Simulations of Abiotic Hydrocarbon Formation in Earth’s Deep Subsurface. Reviews in Mineralogy and Geochemistry, 75, 467-494.

[10]   Charlou, J.L., Donval, J.P., Konn, C., Ondreas, H., Fouquet, Y., Jean Baptiste, P. and Fourré, E. (2010) High Production and Fluxes of H2 and CH4 and Evidence of Abiotic Hydrocarbon Synthesis by Serpentinization in Ultramafic-Hosted Hydrothermal Systems on the Mid-Atlantic Ridge. In: Rona, P., Devey, C., Dyment, J. and Murton, B., Eds., Diversity of Hydrothermal Systems on Slow-Spreading Ocean Ridges, Washington DC, 265-296.

[11]   Proskurowski, G., Lilley, M.D., Seewald, J.S., Fruh-Green, G.L., Olson, E.J., Lupton, J.E., Sylva, S.P. and Kelley, D.S. (2008) Abiogenic Hydrocarbon Production at Lost City Hydrothermal Field. Science, 319, 604-607.

[12]   Fuchida, S., Mizuno, Y., Masuda, H., Toki, T. and Makita, H. (2014) Concentrations and Distributions of Amino Acids in Black and White Smoker Fluids at Temperatures over 200°C. Organic Geochemistry, 66, 98-106.

[13]   Haberstroh, P.R. and Karl, D.M. (1989) Dissolved Free Amino Acids in Hydrothermal Vent Habitats of the Guaymas Basin. Geochimica et Cosmochimica Acta, 53, 2937-2945.

[14]   Horiuchi, T., Takano, Y., Ishibashi, J.I., Marumo, K., Urabe, T. and Kobayashi, K. (2004) Amino Acids in Water Samples from Deep Sea Hydrothermal Vents at Suiyo Seamount, Izu-Bonin Arc, Pacific Ocean. Organic Geochemistry, 35, 1121-1128.

[15]   Klevenz, V., Sumoondur, A., Ostertag-Henning, C. and Koschinsky, A. (2010) Concentrations and Distributions of Dissolved Amino Acids in Fluids from Mid-Atlantic Ridge Hydrothermal Vents. Geochemical Journal, 44, 387-397.

[16]   Lang, S.Q., Früh-Green, G.L., Bernasconi, S.M. and Butterfield, D.A. (2013) Sources of Organic Nitrogen at the Serpentinite-Hosted Lost City Hydrothermal Field. Geobiology, 11, 154-169.

[17]   Svensson, E., Skoog, A. and Amend, J.P. (2004) Concentration and Distribution of Dissolved Amino Acids in a Shallow Hydrothermal System, Vulcano Island (Italy). Organic Geochemistry, 35, 1001-1014.

[18]   Lee, N., Foustoukos, D.I., Sverjensky, D.A., Cody, G.D. and Hazen, R.M. (2014) The Effects of Temperature, pH and Redox State on the Stability of Glutamic Acid in Hydrothermal Fluids. Geochimica et Cosmochimica Acta, 135, 66-86.

[19]   Lee, N., Foustoukos, D.I., Sverjensky, D.A., Hazen, R.M. and Cody, G.D. (2014) Hydrogen Enhances the Stability of Glutamic Acid in Hydrothermal Environments. Chemical Geology, 386, 184-189.

[20]   McCollom, T.M. (2013) The Influence of Minerals on Decomposition of the n-alkyl-α-amino Acid Norvaline under Hydrothermal Conditions. Geochimica et Cosmochimica Acta, 104, 330-357.

[21]   Cox, J.S. and Seward, T.M. (2007) The Reaction Kinetics of Alanine and Glycine under Hydrothermal Conditions. Geochimica et Cosmochimica Acta, 71, 2264-2284.

[22]   Ito, M., Gupta, L.P., Masuda, H. and Kawahata, H. (2006) Thermal Stability of Amino Acids in Seafloor Sediment in Aqueous Solution at High Temperature. Organic Geochemistry, 37, 177-188.

[23]   Orsi, W.D., Edgcomb, V.P., Christman, G.D. and Biddle, J.F. (2013) Gene Expression in the Deep Biosphere. Nature, 499, 205-208.

[24]   Charlou, J.L., Donval, J.P., Fouquet, Y., Jean-Baptiste, P. and Holm, N. (2002) Geochemistry of High H2 and CH4 Vent Fluids Issuing from Ultramafic Rocks at the Rainbow Hydrothermal Field (36°14'N, MAR). Chemical Geology, 191, 345-359.

[25]   Charlou, J.L., Fouquet, Y., Bougault, H., Donval, J.P., Etoubleau, J., Jean-Baptiste, P., Dapoigny, A., Appriou, P. and Rona, P.A. (1998) Intense CH4 Plumes Generated by Serpentinization of Ultramafic Rocks at the Intersection of the 15°20'N Fracture Zone and the Mid-Atlantic Ridge. Geochimica et Cosmochimica Acta, 62, 2323-2333.

[26]   Casella, I.G. and Contursi, M. (2003) Isocratic Ion Chromatographic Determination of Underivatized Amino Acids by Electrochemical Detection. Analytica Chimica Acta, 478, 179-189.

[27]   Yu, H., Ding, Y.S., Mou, S.F., Jandik, P. and Cheng, J. (2002) Simultaneous Determination of Amino Acids and Carbohydrates by Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection. Journal of Chromatography A, 966, 89-97.

[28]   Ozcan, S. and Oenyuva, H.Z. (2006) Improved and Simplified Liquid Chromatography/Atmospheric Pressure Chemical Ionization Mass Spectrometry Method for the Analysis of Underivatized Free Amino Acids in Various Foods. Journal of Chromatography A, 1135, 179-185.

[29]   Yan, D., Li, G., Xiao, X.H., Dong, X.P. and Li, Z.L. (2007) Direct Determination of Fourteen Underivatized Amino Acids from Whitmania Pigra by Using Liquid Chromatography-Evaporative Light Scattering Detection. Journal of Chromatography A, 1138, 301-304.

[30]   Petritis, K., Elfakir, C. and Dreux, M. (2002) A Comparative Study of Commercial Liquid Chromatographic Detectors for the Analysis of Underivatized Amino Acids. Journal of Chromatography A, 961, 9-21.

[31]   Buiarelli, F., Gallo, V., Di Filippo, P., Pomata, D. and Riccardi, C. (2013) Development of a Method for the Analysis of Underivatized Amino Acids by Liquid Chromatography/Tandem Mass Spectrometry: Application on Standard Reference Material 1649a (Urban Dust). Talanta, 115, 966-972.

[32]   Le, A., Ng, A., Kwan, T., Cusmano-Ozog, K. and Cowan, T.M. (2014) A Rapid, Sensitive Method for Quantitative Analysis of Underivatized Amino Acids by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Journal of Chromatography B, 944, 166-174.

[33]   Zhou, G., Pang, H., Tang, Y., Yao, X., Mo, X., Zhu, S., Guo, S., Qian, D., Qian, Y., Su, S., Zhang, L., Jin, C., Qin, Y. and Duan, J.A. (2013) Hydrophilic Interaction Ultra-Performance Liquid Chromatography Coupled with Triple- Quadrupole Tandem Mass Spectrometry for Highly Rapid and Sensitive Analysis of Underivatized Amino Acids in Functional Foods. Amino Acids, 44, 1293-1305.

[34]   Petritis, K., Chaimbault, P., Elfakir, C. and Dreux, M. (2000) Parameter Optimization for the Analysis of Underivatized Protein Amino Acids by Liquid Chromatography and Ionspray Tandem Mass Spectrometry. Journal of Chromatography A, 896, 253-263.

[35]   Piraud, M., Vianey-Saban, C., Petritis, K., Elfakir, C., Steghens, J.P. and Bouchu, D. (2005) Ion-Pairing Reversed- Phase Liquid Chromatography/Electrospray Ionization Mass Spectrometric Analysis of 76 Underivatized Amino Acids of Biological Interest: A New Tool for the Diagnosis of Inherited Disorders of Amino Acid Metabolism. Rapid Communications in Mass Spectrometry, 19, 1587-1602.

[36]   Chaimbault, P., Petritis, K., Elfakir, C. and Dreux, M. (1999) Determination of 20 Underivatized Proteinic Amino Acids by Ion-Pairing Chromatography and Pneumatically Assisted Electrospray Mass Spectrometry. Journal of Chromatography A, 855, 191-202.

[37]   Liu, D.L., Beegle, L.W. and Kanik, I. (2008) Analysis of Underivatized Amino Acids in Geological Samples Using Ion-Pairing Liquid Chromatography and Electrospray Tandem Mass Spectrometry. Astrobiology, 8, 229-241.

[38]   Takano, Y., Chikaraishi, Y. and Ohkouchi, N. (2014) Isolation of Underivatized Amino Acids by Ion-Pair High Performance Liquid Chromatography for Precise Measurement of Nitrogen Isotopic Composition of Amino Acids: Development of Comprehensive LC × GC/C/IRMS Method. International Journal of Mass Spectrometry, In Press.

[39]   Troise, A., Fiore, A., Roviello, G., Monti, S. and Fogliano, V. (2015) Simultaneous Quantification of Amino Acids and Amadori Products in Foods through Ion-Pairing Liquid Chromatography-High-Resolution Mass Spectrometry. Amino Acids, 47, 111-124.

[40]   Armstrong, M., Jonscher, K. and Reisdorph, N.A. (2007) Analysis of 25 Underivatized Amino Acids in Human Plasma Using Ion-Pairing Reversed-Phase Liquid Chromatography/Time-of-Flight Mass Spectrometry. Rapid Communications in Mass Spectrometry, 21, 2717-2726.

[41]   Bar-Nun, A., Bar-Nun, N., Bauer, S.H. and Sagan, C. (1970) Shock Synthesis of Amino Acids in Simulated Primitive Environments. Science, 168, 470-472.

[42]   Hennet, R.J.C., Holm, N.G. and Engel, M.H. (1992) Abiotic Synthesis of Amino Acids under Hydrothermal Conditions and the Origin of Life: A Perpetual Phenomenon? Naturwissenschaften, 79, 361-365.

[43]   Marshall, W.L. (1994) Hydrothermal Synthesis of Amino Acids. Geochimica et Cosmochimica Acta, 58, 2099-2106.

[44]   Yanagawa, H. and Kobayashi, K. (1992) Chapter 8. An Experimental Approach to Chemical Evolution in Submarine Hydrothermal Systems. Origins of Life and Evolution of the Biosphere, 22, 147-159.

[45]   Andersson, E. and Holm, N.G. (2000) The Stability of Some Selected Amino Acids under Attempted Redox Constrained Hydrothermal Conditions. Origins of Life and Evolution of the Biosphere, 30, 9-23.

[46]   Islam, M.N., Kaneko, T. and Kobayashi, K. (2001) Determination of Amino Acids Formed in a Supercritical Water Flow Reactor Simulating Submarine Hydrothermal Systems. Analytical Sciences, 17, i1631-i1634.

[47]   Managau, L.M. and Courtot, P. (1987) Dissolved Free Amino Acids of Coastal Seawater (Strait of Brest, France). Journal de Recherche Oceanographique, 12, 39-42.

[48]   Henrichs, S.M., Farrington, J.W. and Lee, C. (1984) Peru Upwelling Region Sediments near 15°S. 2. Dissolved Free and Total Hydrolyzable Amino Acids. Limnology and Oceanography, 29, 20-34.

[49]   Waters-Corporation (2009) ACQUITY UPLC® Systems—Overcome the Challenges of Analytical Laboratories.

[50]   Hiraoka, K., Murata, K. and Kudaka, I. (1995) Do the Electrospray Mass Spectra Reflect the Ion Concentrations in Sample Solution? Journal of the Mass Spectrometry Society of Japan, 43, 127-138.

[51]   Houghton, R. and Grace, P. (2008) UHPLC—Why All the Hype? Chromatography Today, 5-7.

[52]   Waters-Corporation (2004) New Quadrupole MS Detection Capabilities for Ultra Performance LC.

[53]   Waters-Corporation (2007) An Introduction to UPLC® Technology: Improve Productivity and Data Quality.

[54]   Sommerville, K. and Preston, T. (2001) Characterisation of Dissolved Combined Amino Acids in Marine Waters. Rapid Communications in Mass Spectrometry, 15, 1287-1290.

[55]   Daumas, R.A. (1976) Variations of Particulate Proteins and Dissolved Amino Acids in Coastal Seawater. Marine Chemistry, 4, 225-242.

[56]   Farkas, E. and Sóvágó, I. (2002) Amino Acids, Peptides and Proteins. The Royal Society of Chemistry, London.

[57]   Yamauchi, O. and Odani, A. (1996) Stability Constants of Metal Complexes of Amino Acids with Charged Side Chains—Part I: Positively Charged Side Chains. Pure and Applied Chemistry, 68, 469-496.

[58]   Evans, G.W. (1973) Copper Homeostasis in the Mammalian System. Physiological Reviews, 53, 535-570.

[59]   Gupta, A., Loew, G.H. and Lawless, J. (1983) Interaction of Metal Ions and Amino Acids: Possible Mechanisms for the Adsorption of Amino Acids on Homoionic Smectite Clays. Inorganic Chemistry, 22, 111-120.

[60]   Hedges, J.I. and Hare, P.E. (1987) Amino Acid Adsorption by Clay Minerals in Distilled Water. Geochimica et Cosmochimica Acta, 51, 255-259.

[61]   Zaia, D.A.M., Vieira, H.J. and Zaia, C.T.B.V. (2002) Adsorption of L-Amino Acids on Sea Sand. Journal of the Brazilian Chemical Society, 13, 679-681.

[62]   Henrichs, S.M. and Sugai, S.F. (1993) Adsorption of Amino Acids and Glucose by Sediments of Resurrection Bay, Alaska, USA: Functional Group Effects. Geochimica et Cosmochimica Acta, 57, 823-835.

[63]   Zaia, D.A.M. (2004) A Review of Adsorption of Amino Acids on Minerals: Was It Important for Origin of Life? Amino Acids, 27, 113-118.

[64]   Holm, N.G., Dowler, M.J., Wadsten, T. and Arrhenius, G. (1983) β-FeOOH·Cln (akaganéite) and Fe1-xO (wüstite) in Hot Brine from the Atlantis II Deep (Red Sea) and the Uptake of Amino Acids by Synthetic β-FeOOH·Cln. Geochimica et Cosmochimica Acta, 47, 1465-1470.

[65]   Hazen, R.M. and Sverjensky, D.A. (2010) Mineral Surfaces, Geochemical Complexities, and the Origins of Life. Cold Spring Harbor Perspectives in Biology, 2.

[66]   Lahav, N. and Chang, S. (1976) The Possible Role of Solid Surface Area in Condensation Reactions during Chemical Evolution: Reevaluation. Journal of Molecular Evolution, 8, 357-380.

[67]   Lambert, J.F. (2008) Adsorption and Polymerization of Amino Acids on Mineral Surfaces: A Review. Origins of Life and Evolution of Biospheres, 38, 211-242.

[68]   Kaiser, K. and Benner, R. (2009) Biochemical Composition and Size Distribution of Organic Matter at the Pacific and Atlantic Time-Series Stations. Marine Chemistry, 113, 63-77.