By Michail A. Alterman, Peter Hunziker
Amino Acid research (AAA) is a vital part of analytical biochemistry. In a comparatively couple of minutes, the diversity of AAA tools has developed dramatically with extra equipment transferring to using mass spectrometry (MS) as a detection strategy. one other new point is miniaturization. even if, most significantly, AAA at the present time might be considered within the context of Metabolomics as part of platforms Biology. Amino Acid research: tools and Protocols offers a wide spectrum of all to be had equipment making an allowance for readers to decide on the tactic that almost all matches their specific laboratory set-up and analytical wishes. during this quantity, a reader can locate chapters describing common in addition to particular techniques to the pattern training. a couple of chapters describe particular functions of AAA in medical chemistry in addition to in nutrition research, microbiology, marine biology, drug metabolism, even archeology. Separate chapters are dedicated to the appliance of AAA for protein quantitation and chiral AAA. Written within the hugely profitable equipment in Molecular Biology™ sequence layout, chapters include introductions to their respective themes, lists of the required fabrics and reagents, step by step, effortlessly reproducible laboratory protocols, and notes on troubleshooting and heading off identified pitfalls. Authoritative and obtainable, Amino Acid research: tools and Protocols offers the most important recommendations that may be utilized throughout a number of disciplines by means of a person concerned about biomedical study or existence sciences.
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Additional resources for Amino Acid Analysis: Methods and Protocols (Methods in Molecular Biology, v828)
Savant ISS110 SpeedVac Concentrator (ThermoFisher Scientific/Thermo Electron LED GmbH, Vienna, Austria). 4. Glass inserts (VWR Österreich, Vienna, Austria). 3. Methods The HILIC-MS/MS method as well as the enrichment procedure was recently published and has been modified for improvement (1). 1. Liquid Chromatography Flow rate was set to 500 μL/min, autosampler temperature to 5°C, and injection volume to 10 μL. Gradient elution was performed using Eluent A (98% v/v water, 1% v/v ACN, 1% v/v formic acid ) and Eluent B (98% v/v ACN, 1% v/v water, 1% formic acid) as follows: starting conditions of 90% B were held for 1 min, then B was decreased to 10% within 7 min and was constant for 1 min.
Notes 1. Certain commercial equipment, instruments, or materials are identified in this chapter to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose. 2. Solutions containing trifluoroacetic acid have limited stability and the breakdown products can increase the chemical background seen in mass spectrometry.
Callahan DL et al (2007) Relationships of nicotianamine and other amino acids with nickel, zinc and iron in Thlaspi hyperaccumulators. New Phytol 176: 836–848 9. Tag ultra performance liquid chromatography-electrospray ionization-MS/MS with multiple reaction monitoring. Anal Chem 82: 548–558 10. Bosch L, Alegria A, and Farre R (2006) Application of the 6-aminoquinolyl-Nhydroxysccinimidyl carbamate (AQC) reagent to the RP-HPLC determination of amino acids in infant foods. J Chromatogr B Analyt Technol Biomed Life Sci 831: 176–183 11.