M E T H O D S I N M O L E C U L A R B I O L O G Y ™

Series EditorJohn M. Walker

School of Life SciencesUniversity of Hertfordshire

Hatfield, Hertfordshire, AL10 9AB, UK

For further volumes: http://www.springer.com/series/7651

Amino Acid Analysis

Methods and Protocols

Edited by

Michail A. Alterman

Tumor Vaccines and Biotechnology Branch, CBER, FDA, Bethesda, MD, USA

Peter Hunziker

Functional Genomics Center, University of Zűrich, Zűrich, Switzerland

ISSN 1064-3745 e-ISSN 1940-6029ISBN 978-1-61779-444-5 e-ISBN 978-1-61779-445-2DOI 10.1007/978-1-61779-445-2Springer New York Dordrecht Heidelberg London

Library of Congress Control Number: 2011941583

© Springer Science+Business Media, LLC 2012All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden.The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights.

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Humana Press is part of Springer Science+Business Media (www.springer.com)

EditorsMichail A. AltermanTumor Vaccines and Biotechnology BranchCBER, FDABethesda, MD, USAMichail.Alterman@fda.hhs.gov

Peter HunzikerFunctional Genomics CenterUniversity of ZürichZürich, Switzerlandpeter.hunziker@access.uzh.ch

v

Preface

Amino Acid Analysis (AAA) has been an integral part of analytical biochemistry for almost 60 years. AAA was originally developed by Moore and Stein and was at the very heart of their work on the mechanism of enzyme catalysis for which they were awarded a Nobel Prize in Chemistry in 1972. In a relatively short time since the previous AAA book in this series has been published (10 years), the variety of AAA methods changed dramatically with more methods shifting to the use of mass spectrometry (MS) as a detection method. At the same time, a number of old techniques acquired a new make-up, like combination of AccQ-Tag with UPLC and MS, instead of HPLC and fl uorescence. Another new aspect is miniaturization. One of the chapters in this book describes an AAA in a single cell. However, the most important aspect is that AAA in this day and age should be viewed in the context of Metabolomics as a part of Systems Biology.

Historically, analysis of amino acids (AA) includes derivatization (pre, on-, or post-column) coupled with chromatographic separation. A wide variety of separation techniques were applied to separation of AA over the years. This list includes ion-exchange LC, reverse-phase HPLC, gas chromatography (GC), and capillary electrophoresis (CE). Recent advances in mass spectrometry (MS) led to the application of electrospray ionization cou-pled with LC or CE for AA detection which may also allow the analysis of underivatized AA. Two most recent technological advances in AAA include the application of MALDI TOF MS and TOF/TOF MS/MS and microfl uidics. AAA techniques evolve and follow the bioanalytical technological advances.

Sample preparation for AAA plays a critical role in the successful implementation of AAA. Correspondingly, in this book a reader can fi nd chapters describing general as well as specifi c approaches to the sample preparation. A number of chapters describe different applications of AAA. Some chapters describe specifi c applications of AAA in clinical chem-istry as well as in food analysis, microbiology, and other biomedical fi elds. Separate chapters are devoted to the application of AAA for protein quantitation and chiral AAA.

Our goal was to present a spectrum of all available methods for readers to choose the method that most suits particular laboratory and needs. And, at the same time we attempted to present more than one method for each application or detection/separation approach so that again the readers can fi nd the one that most suits their needs and available laboratory conditions.

What is unique about this book, and in essence about AAA itself, is that it is related and is of interest to anyone involved in biomedical research or, in general, in life sciences. One can fi nd here techniques essential in medicine, or in drug metabolism, or cell biology, even in archeology, in meat industry, in marine biology, in agriculture, and the list goes on. All of the described techniques are multifaceted and in many cases can serve as a blueprint for the analysis of other chemically related classes of metabolites.

Bethesda, MD, USA Michail A. AltermanZürich, Switzerland Peter Hunziker

vii

Contents

Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vContributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

1 Rapid LC–MS/MS Profiling of Protein Amino Acids and Metabolically Related Compounds for Large-Scale Assessment of Metabolic Phenotypes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1Liping Gu, A. Daniel Jones, and Robert L. Last

2 Combination of an AccQ·Tag-Ultra Performance Liquid Chromatographic Method with Tandem Mass Spectrometry for the Analysis of Amino Acids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13Carolina Salazar, Jenny M. Armenta, Diego F. Cortés,and Vladimir Shulaev

3 Isotope Dilution Liquid Chromatography-Tandem Mass Spectrometry for Quantitative Amino Acid Analysis. . . . . . . . . . . . . . . . . . . . . 29David M. Bunk and Mark S. Lowenthal

4 Analysis of Underivatized Amino Acids: Zwitterionic Hydrophilic Interaction Chromatography Combined with Triple Quadrupole Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39Madeleine Dell’mour, Gunda Koellensperger, and Stephan Hann

5 Amino Acid Analysis via LC–MS Method After Derivatization with Quaternary Phosphonium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47Shinsuke Inagaki and Toshimasa Toyo’oka

6 Amino Acid Analysis by Hydrophilic Interaction Chromatography Coupled with Isotope Dilution Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . 55Megumi Kato and Akiko Takatsu

7 A Universal HPLC-MS Method to Determine the Stereochemistry of Common and Unusual Amino Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63Sonja Hess

8 Amino Acid Analysis by Capillary Electrophoresis-Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77Akiyoshi Hirayama and Tomoyoshi Soga

9 New Advances in Amino Acid Profiling by Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry . . . . . . . . . . . . . . 83Philip Britz-McKibbin

10 Optimal Conditions for the Direct RP-HPLC Determination of Underivatized Amino Acids with Online Multiple Detection . . . . . . . . . . . . 101A. Pappa-Louisi, P. Agrafiotou, and S. Sotiropoulos

viii Contents

11 Absolute Quantitation of Proteins by Acid Hydrolysis Combined with Amino Acid Detection by Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . 115Olga A. Mirgorodskaya, Roman Körner, Yuri P. Kozmin, and Peter Roepstorff

12 Amino Acid Analysis by Means of MALDI TOF Mass Spectrometry or MALDI TOF/TOF Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121Natalia V. Gogichaeva and Michail A. Alterman

13 Heptafluorobutyl Chloroformate-Based Sample Preparation Protocol for Chiral and Nonchiral Amino Acid Analysis by Gas Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137Petr Šimek, Petr Hušek, and Helena Zahradníčková

14 The EZ:Faast Family of Amino Acid Analysis Kits: Application of the GC-FID Kit for Rapid Determination of Plasma Tryptophan and Other Amino Acids . . . . . . . . . . . . . . . . . . . . . . . . 153Abdulla A.-B. Badawy

15 Amino Acid Analysis in Physiological Samples by GC–MS with Propyl Chloroformate Derivatization and iTRAQ–LC–MS/MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165Katja Dettmer, Axel P. Stevens, Stephan R. Fagerer, Hannelore Kaspar, and Peter J. Oefner

16 Automated Analysis of Primary Amino Acids in Plasma by High-Performance Liquid Chromatography . . . . . . . . . . . . . . . . . . . . . . . . 183Durk Fekkes

17 RP-LC of Phenylthiocarbamyl Amino Acid Adducts in Plasma Acetonitrile Extracts: Use of Multiple Internal Standards and Variable Wavelength UV Detection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201Lionella Palego, Gino Giannaccini, and Antonio Lucacchini

18 Quantification of Underivatised Amino Acids on Dry Blood Spot, Plasma, and Urine by HPLC–ESI–MS/MS. . . . . . . . . . . . . . . . . . . . . . . . . . . 219Giuseppe Giordano, Iole Maria Di Gangi, Antonina Gucciardi,and Mauro Naturale

19 Capillary Electrophoresis of Free Amino Acids in Physiological Fluids Without Derivatization Employing Direct or Indirect Absorbance Detection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243Gordana D. Žunić, Slavica Spasić, and Zorana Jelić-Ivanović

20 Measurement of 3-Nitro-Tyrosine in Human Plasma and Urine by Gas Chromatography-Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . 255Dimitrios Tsikas, Anja Mitschke, and Frank-Mathias Gutzki

21 Analysis of Hydroxyproline in Collagen Hydrolysates . . . . . . . . . . . . . . . . . . . 271Tobias Langrock and Ralf Hofmann

22 Innovative and Rapid Procedure for 4-Hydroxyproline Determination in Meat-Based Foods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 281Maria Cristina Messia and Emanuele Marconi

ixContents

23 Multiple Reaction Monitoring for the Accurate Quantification of Amino Acids: Using Hydroxyproline to Estimate Collagen Content . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291Michelle L. Colgrave, Peter G. Allingham, Kerri Tyrrell, and Alun Jones

24 Sequential Injection Chromatography for Fluorimetric Determination of Intracellular Amino Acids in Marine Microalgae. . . . . . . . . . 305Marilda Rigobello-Masini and Jorge C. Masini

25 Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS-MS. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 317Björn Thiele, Nadine Stein, Marco Oldiges, and Diana Hofmann

26 Wheat Gluten Amino Acid Analysis by High-Performance Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 329Ine Rombouts, Bert Lagrain, Lieve Lamberts, Inge Celus, Kristof Brijs, and Jan A. Delcour

27 Preparative HPLC Separation of Underivatized Amino Acids for Isotopic Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339Jennifer A. Tripp and James S.O. McCullagh

28 Quantification of Amino Acids in a Single Cell by Microchip Electrophoresis with Chemiluminescence Detection . . . . . . . . . . . . . . . . . . . . 351Yi-Ming Liu and Shulin Zhao

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359

xi

Contributors

P. AGRAFIOTOU • Laboratory of Physical Chemistry, Department of Chemistry , Aristotle University of Thessaloniki , Thessaloniki , Greece

PETER G. ALLINGHAM • Cooperative Research Centre for Sheep Industry Innovation, CJ Hawkins Homestead, University of New England , Armidale , NSW , Australia

MICHAIL A. ALTERMAN • Tumor Vaccines and Biotechnology Branch , CBER, FDA , Bethesda , MD , USA

JENNY M. ARMENTA • Waters Corporation , Beverly , MA , USA ABDULLA A.-B. BADAWY • The Cardiff School of Health Sciences, University of Wales

Institute Cardiff (UWIC) , Western Avenue, Cardiff, Wales , UK KRISTOF BRIJS • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science

and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

PHILIP BRITZ-MCKIBBIN • Department of Chemistry and Chemical Biology , McMaster University , Hamilton , ON , Canada

DAVID M. BUNK • Analytical Chemistry Division , National Institute of Standards and Technology , Gaithersburg , MD , USA

INGE CELUS • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

MICHELLE L. COLGRAVE • CSIRO Division of Livestock Industries, Queensland Bioscience Precinct , St Lucia , QLD , Australia

DIEGO F. CORTÉS • Virginia Bioinformatics Institute, Virginia Polytechnic Institute and State University , Blacksburg , VA , USA

JAN A. DELCOUR • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

MADELEINE DELL’MOUR • Department of Chemistry , University of Natural Resources and Life Sciences – BOKU Vienna , Vienna , Austria

KATJA DETTMER • Institute of Functional Genomics, University of Regensburg , Regensburg , Germany

STEPHAN R. FAGERER • Department of Chemistry and Applied Biosciences, Laboratory of Organic Chemistry , ETH Zürich , Zürich , Switzerland

DURK FEKKES • Department of Psychiatry, Erasmus MC , University Medical Center Rotterdam , Rotterdam , Netherlands

IOLE MARIA DI GANGI • Department of Pediatrics , University of Padova , Padova , Italy GINO GIANNACCINI • Department of Psychiatry, Neurobiology, Pharmacology

and Biotechnology , University of Pisa , Pisa , Italy GIUSEPPE GIORDANO • Department of Pediatrics , University of Padova , Padova , Italy

xii Contributors

NATALIA V. GOGICHAEVA • Analytical Proteomics Laboratory , University of Kansas , Lawrence , KS , USA

LIPING GU • Department of Biology and Microbiology , South Dakota State University , Brookings , SD , USA

ANTONINA GUCCIARDI • Department of Pediatrics , University of Padua , Padova , Italy FRANK-MATHIAS GUTZKI • Institute of Clinical Pharmacology,

Hannover Medical School , Hannover , Germany STEPHAN HANN • Department of Chemistry , University of Natural Resources

and Life Sciences – BOKU Vienna , Vienna , Austria SONJA HESS • Proteome Exploration Laboratory , California Institute of Technology ,

BI 211 , MC139-74 , Pasadena , CA , USA AKIYOSHI HIRAYAMA • Institute for Advanced Biosciences, Keio University ,

Kakuganji, Tsuruoka, Yamagata , Japan RALF HOFFMANN • Institute of Bioanalytical Chemistry, Faculty of Chemistry

and Mineralogy and Center for Biotechnology and Biomedicine, Universität Leipzig , Leipzig , Germany

DIANA HOFMANN • Central Division of Analytical Chemistry/BioSpec , Jülich , Germany PETR HUŠEK • Laboratory of Analytical Biochemistry , Biology Centre,

Academy of Sciences of the Czech Republic , České Budějovice , Czech Republic SHINSUKE INAGAKI • Laboratory of Analytical and Bio-Analytical Chemistry ,

School of Pharmaceutical Sciences, and Global COE Program, University of Shizuoka , Shizuoka , Japan

ZORANA JELIĆ-IVANOVIĆ • Department of Medical Biochemistry, Faculty of Pharmacy , University of Belgrade , Belgrade , Serbia

A. DANIEL JONES • Department of Biochemistry and Molecular Biology, and Department of Chemistry , Michigan State University , East Lansing , MI , USA

ALUN JONES • The University of Queensland, Institute for Molecular Bioscience , Brisbane , QLD , Australia

HANNELORE KASPAR • Institute of Functional Genomics, University of Regensburg , Regensburg , Germany

MEGUMI KATO • Bio-Medical Standard Section , National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST) , Tsukuba, Ibaraki , Japan

GUNDA KOELLENSPERGER • Department of Chemistry , University of Natural Resources and Life Sciences – BOKU Vienna , Vienna , Austria

ROMAN KÖRNER • Max-Planck Institute of Biochemistry , Martinsried , Germany YURI P. KOZMIN • Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS. Ul.

Miklukho-Maklaya , GSP Moscow , Russia BERT LAGRAIN • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science

and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

LIEVE LAMBERTS • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

TOBIAS LANGROCK • Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy and Center for Biotechnology and Biomedicine, Universität Leipzig , Leipzig , Germany

xiiiContributors

ROBERT L. LAST • Departments of Biochemistry and Molecular Biology and Department of Plant Biology , Michigan State University , East Lansing , MI , USA

YI-MING LIU • Department of Chemistry and Biochemistry , Jackson State University , Jackson , MS , USA

MARK S. LOWENTHAL • Analytical Chemistry Division , National Institute of Standards and Technology , Gaithersburg , MD , USA

ANTONIO LUCACCHINI • Department of Psychiatry, Neurobiology, Pharmacology and Biotechnology , University of Pisa , Pisa , Italy

EMANUELE MARCONI • DISTAAM, Università degli Studi del Molise , Campobasso , Italy

JORGE C. MASINI • Instituto de Química, Universidade de São Paulo , São Paulo , Brazil JAMES S. O. MCCULLAGH • Department of Chemistry , University of Oxford ,

Oxford , UK MARIA CRISTINA MESSIA • DISTAAM, Università degli Studi del Molise ,

Campobasso , Italy OLGA A. MIRGORODSKAYA • Research Institute of Infl uenza , Saint-Petersburg , Russia ANJA MITSCHKE • Institute of Clinical Pharmacology, Hannover Medical School ,

Hannover , Germany MAURO NATURALE • Department of Pediatrics , University of Padova , Padova , Italy PETER J. OEFNER • Institute of Functional Genomics, University of Regensburg ,

Regensburg , Germany MARCO OLDIGES • IBG-1: Biotechnology, Forschungszentrum Jülich GmbH ,

Jülich , Germany LIONELLA PALEGO • Department of Psychiatry, Neurobiology, Pharmacology

and Biotechnology , University of Pisa , Pisa , Italy A. PAPPA-LOUISI • Laboratory of Physical Chemistry, Department of Chemistry ,

Aristotle University of Thessaloniki , Thessaloniki , Greece MARILDA RIGOBELLO-MASINI • Instituto de Química, Universidade de São Paulo ,

São Paulo , Brazil PETER ROEPSTORFF • Department of Biochemistry and Molecular Biology ,

University of Southern Denmark , Odense M , Denmark INE ROMBOUTS • Laboratory of Food Chemistry and Biochemistry , Leuven Food Science

and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven , Leuven , Belgium

CAROLINA SALAZAR • Department of Biological Sciences , University of North Texas,College of Arts and Sciences , Denton , TX , USA

VLADIMIR SHULAEV • Department of Biological Sciences , University of North Texas,College of Arts and Sciences , Denton , TX , USA

PETR ŠIMEK • Laboratory of Analytical Biochemistry , Biology Centre, Academy of Sciences of the Czech Republic , České Budějovice , Czech Republic

S. SOTIROPOULOS • Laboratory of Physical Chemistry, Department of Chemistry , Aristotle University of Thessaloniki , Thessaloniki , Greece

TOMOYOSHI SOGA • Institute for Advanced Biosciences, Keio University , Kakuganji, Tsuruoka, Yamagata , Japan

SLAVICA SPASIĆ • Department of Medical Biochemistry, Faculty of Pharmacy , University of Belgrade , Belgrade , Serbia

NADINE STEIN • IBG-1: Biotechnology , Jülich , Germany

xiv Contributors

AXEL P. STEVENS • Institute of Functional Genomics, University of Regensburg , Regensburg , Germany

AKIKO TAKATSU • Bio-Medical Standard Section , National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST) , Tsukuba, Ibaraki , Japan

BJÖRN THIELE • Institute of Bio- and Geosciences, Forschungszentrum Jülich GmbH, ICG-3 , Jülich , Germany

TOSHIMASA TOYO’OKA • Laboratory of Analytical and Bio-Analytical Chemistry , School of Pharmaceutical Sciences, and Global COE Program, University of Shizuoka , Surugaku, Shizuoka , Japan

JENNIFER A. TRIPP • Department of Chemistry and Biochemistry , San Francisco State University , San Francisco , CA , USA

DIMITRIOS TSIKAS • Institute of Clinical Pharmacology, Hannover Medical School , Hannover , Germany

KERRI TYRRELL • CSIRO Division of Livestock Industries, Queensland Bioscience Precinct , St Lucia , QLD , Australia

HELENA ZAHRADNÍČKOVÁ • Laboratory of Analytical Biochemistry , Biology Centre, Academy of Sciences of the Czech Republic , České Budějovice , Czech Republic

SHULIN ZHAO • Department of Chemistry and Biochemistry , Jackson State University , Jackson , MS , USA

GORDANA D. ŽUNIĆ • Institute for Medical Research, Military Medical Academy , Belgrade , Serbia