**Meet the editors**

Ana Varela Coelho received her Ph.D. in 1998 from the University of Évora, Portugal, where she has taught several biochemistry curricular units for thirteen years. She was an Invited Professor at the Instituto de Tecnologia Química e Biológica – Universidade Nova de Lisboa, Oeiras, Portugal and then joined this Institute as an assistant researcher where she coordinates the Mass

Spectrometry Laboratory since 2002. She has published more than eighty research articles in peer reviewed journal and participated in more than forty scientific projects. Her research interests include the use of tandem mass spectrometry in proteomic, peptidomic and metabolomic applications, in particular in the study of biological processes with biomedical applications, namely in tissue and organ regeneration.

Catarina F. Franco concluded her Ph.D. in Biochemistry at New University of Lisbon in 2011. During this period Catarina studied the differential protein expression during arm tip regeneration events of starfish specie using proteomic and mass spectrometry based approaches. In 2012, Catarina started a post-doctoral fellow at Instituto de Tecnologia Química e Biológica. Currently she is in-

terested in using tandem mass spectrometry tools for the characterization of protein post-translational modifications occurring during specific stages of regeneration of the starfish radial nerve cords.

Contents

**Preface VII**

**to Control these Effects 3**

Diogo Ribeiro Demartini

**Antimalarial Agents 61**

Lamosa and Ana V. Coelho

Maria Rosário Bronze

Chapter 1 **Matrix Effects in Mass Spectrometry Combined with Separation**

Luigi Silvestro, Isabela Tarcomnicu and Simona Rizea Savu

Chapter 2 **A Short Overview of the Components in Mass Spectrometry Instrumentation for Proteomics Analyses 39**

**Section 2 Molecular Characterization by Mass Spectrometry 59**

Chapter 4 **Mass Spectrometry Strategies for Structural Analysis of Carbohydrates and Glycoconjugates 81** Guilherme L. Sassaki and Lauro Mera de Souza

Chapter 5 **Tandem MS and NMR: An Efficient Couple for the Characterization of Saponins 117**

Chapter 3 **Contribution of Mass Spectrometry to the Study of**

**Methods — Comparison HPLC, GC and Discussion on Methods**

Ana Raquel Sitoe, Francisca Lopes, Rui Moreira, Ana Coelho and

Rita Laires, Kamila Koci, Elisabete Pires, Catarina Franco, Pedro

Chapter 6 **Post-Translational Modification Profiling of Burn-Induced — Insulin Resistance and Muscle Wasting 137**

Xiao-Ming Lu, Ronald G. Tompkins and Alan J. Fischman

**Section 1 General Aspects 1**

## Contents

### **Preface XI**



### **Section 2 Molecular Characterization by Mass Spectrometry 59**


Preface

odology.

Tandem mass spectrometry started as a tool for the structural characterization of simple mo‐ lecular components, presently, this analytical approach is being extensively used for the characterization and quantification of small molecules, and also of complex biomolecules. The expeditious evolution at the instrumentation level and the increasing number of new applications in a variety of fields indicates that a lot should still be expected from this meth‐

This book comprises six chapters divided in two sections, which provides a current review of some general aspects in mass spectrometry and various applications of tandem mass spectrometry in the structural characterization of a diversity of compounds, complementing the previous books in this Tandem Mass Spectrometry series. In the first chapter are descri‐ bed several methods to overcome the matrix effects associated with mass spectrometry and its hyphenated chromatographic methods. The second chapter provides a description of the instrumental components relevant for proteomic applications. The second section includes four chapters on the characterization of antimalarial agents, carbohydrates, glycoconjugates,

Thanks are expressed to the contributing authors who provided their competence and lead‐ ership in the specific field attempting to provide comprehensive reviews. Finally, I would like to thank Ms. Iva Simcic, Publishing Process Manager, and other InTech publication

> **Ana Varela Coelho and Catarina de Matos Ferraz Franco** Institute of Technology, Chemistry and Biology (ITQB)

> > Mass Spectrometry Lab

Portugal

saponines and on the profiling of post-translational modifications.

staffs for their patience and professional support.

## Preface

Tandem mass spectrometry started as a tool for the structural characterization of simple mo‐ lecular components, presently, this analytical approach is being extensively used for the characterization and quantification of small molecules, and also of complex biomolecules.

The expeditious evolution at the instrumentation level and the increasing number of new applications in a variety of fields indicates that a lot should still be expected from this meth‐ odology.

This book comprises six chapters divided in two sections, which provides a current review of some general aspects in mass spectrometry and various applications of tandem mass spectrometry in the structural characterization of a diversity of compounds, complementing the previous books in this Tandem Mass Spectrometry series. In the first chapter are descri‐ bed several methods to overcome the matrix effects associated with mass spectrometry and its hyphenated chromatographic methods. The second chapter provides a description of the instrumental components relevant for proteomic applications. The second section includes four chapters on the characterization of antimalarial agents, carbohydrates, glycoconjugates, saponines and on the profiling of post-translational modifications.

Thanks are expressed to the contributing authors who provided their competence and lead‐ ership in the specific field attempting to provide comprehensive reviews. Finally, I would like to thank Ms. Iva Simcic, Publishing Process Manager, and other InTech publication staffs for their patience and professional support.

### **Ana Varela Coelho and Catarina de Matos Ferraz Franco**

Institute of Technology, Chemistry and Biology (ITQB) Mass Spectrometry Lab Portugal

**Section 1**

**General Aspects**

**Section 1**

**General Aspects**

**Chapter 1**

**Matrix Effects in Mass Spectrometry Combined with**

Even in the early times of chromatography with conventional detectors (i.e. UV/VIS, FID) it became evident that different sample matrices present peculiar interfering compounds, and the importance of using appropriate spiked matrix calibrators in order to get reliable quanti‐ tative results was recognized. In these conditions, however, the main concern was the presence of coeluting compounds giving similar detector responses, while the risk to alter the detector

Coupling liquid chromatography with mass spectrometry (LC-MS) was an important step forward because polar and thermally unstable compounds could be effectively analyzed and the poor specificity of previous detectors was overcome. The main steps to the hyphenation of the two separation techniques were made by Doles and Fenn with the development of the atmospheric pressure ionization (API) interfaces (Doles et al, 1968; Whitehouse et al, 1985; Fenn et al, 1989; Mallet et al, 2004). In short time LC-MS/MS has become an important tool for the analysis of drugs and metabolites from biological fluids, or for trace analysis from complex mixtures with many applications, e.g. pharmacokinetic studies of pharmaceuticals or the study of proteomics. John Fenn received in 2002 the Nobel Prize in Chemistry for his contribution to

This huge improvement in selectivity brought quickly to a simplification of separation methods and/or sample preparation but on the other hand unexpected quantitative or even

> © 2013 Silvestro et al.; licensee InTech. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2013 Silvestro et al.; licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**Separation Methods — Comparison HPLC, GC and**

**Discussion on Methods to Control these Effects**

Luigi Silvestro, Isabela Tarcomnicu and

**1.1. The evolution of the concept of matrix**

response of the analyte was not yet an issue.

the development of the electrospray ionization (ESI) technique.

Additional information is available at the end of the chapter

Simona Rizea Savu

**1. Introduction**

http://dx.doi.org/10.5772/55982
