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X‐ray fluorescence (XRF) spectroscopy is a well‐established analytical technique being used extensively for mining, metallurgy, petroleum, and geological studies, though not widely used for biological applications. During the past decade, XRF spectrometry has gone through major changes in the field of biological sciences. This book is a guide which provides an up‐to‐date review of XRF spectrometry for biological, medical, food, environmental, and plant science researchers. It covers the basic principles and latest developments in instrumentation and applications of X‐ray fluorescence in biological sciences. This also provides a thoroughly updated and expanded overview to industry professionals in X‐ray analysis over the last decades. The main feature of this book is that it provides information about XRF techniques and procedures for qualitative and quantitative analysis of biological specimens worth modern applications and industrial trends.

The chapters are contributed by independent groups in the world. Four chapters are contributed by the members of editorial advisory board of the journal “X‐Ray Spectrometry” from Wiley. The chapters are divided into six parts. Part 1 is a general introduction of XRF. Parts 2 and 3 are most advanced methods of SR‐XRF and TXRF, which are micro‐XRF, high sensitivity (low detection limit) XRF. Part 4, a beginner’s guide, is one of the characteristics of the present book. Parts 5 and 6 are the main parts of the present book.

Part I (General Introduction) consists of seven (07) chapters. Chapter 1 describes about the XRF and comparison with other analytical methods such as AAS, ICP‐AES, LA‐ICP‐MS, IC, LIBS, SEM‐EDS, and XRD. Chapter 2 highlights the significant role of different XRF configurations for both multi‐elemental bulk analysis and element distribution within vegetal tissues. In Chapter 3, the application of XRF analysis is described for the chemical compositions of tea and coffee samples. Chapter 4 deals with total reflection X‐ray fluorescence (TXRF) spectrometry and its suitability for biological samples. In this chapter, the fundamentals, basic principles, and theoretical aspects of TXRF have been discussed along with its advantages and limitations. Chapter 5 describes the use and application of μ‐XRF and XANES to understand the interaction process and mechanism between microorganisms and heavy metal. Chapter 6 covers the details of EDXRF techniques for the application to clinical samples such as blood and hair. In Chapter 7, all considerations related with the sample preparation process are summarized which is very crucial for XRF analysis.

Part II (Synchrotron Radiation XRF) consists of five (05) chapters which show how Synchrotron Radiation XRF (SRXRF) can be used to provide analytical information in biological sciences for elemental composition. Chapter 8 covers numerous aspects of SRXRF and its applications. This chapter highlights the usefulness of XRF technique for the elemental characterization of different sample matrixes in non‐destructive manners. Chapter 9 deals with the application of SR‐based micro‐XRF spectroscopy for plants. Chapter 10 covers the application of μ‐XRF to study toxic elements in plants. Chapter 11 highlights the application of micro‐XRF for the analysis of benthonic fauna (earthworm and nematodes) in soils and sediments. Chapter 12 discusses in detail the use of SRXRF for the analysis of microelements in biopsy tissues.

Part III (Total Reflection XRF) consists of four (04) chapters which describe in detail the principles and basic fundamentals of total reflection XRF (TXRF) along with their biological applications. Chapter 13 covers the applications of TXRF for the trace element determinations in marine organisms, blood samples, saliva and oral fluids, hairs, nails, kidney stones, urine samples, and forensic samples. Chapter 14 demonstrates the applications of combined X‐ray reflectivity ( XRR ) and grazing incidence X‐ray fluorescence (GIXRF) technique for the characterization of thin films and nano‐structured materials. Chapter 15 deals with the analysis of alcoholic and non‐alcoholic beverages by TXRF. Chapter 16 describes the details of using TXRF and XRT techniques for trace elemental analysis of blood and serum samples.

Part IV (Beginner's Guide) consists of four (04) chapters which cover the basics theory of XRF and historical fundamentals of XRF instruments, quantitative analysis methods, electronics and instrumentation, methods of using XRF to study biological samples. Chapter 17 introduces the atomic physics of the XRF spectrometry which is very useful for beginners to learn for its applications. Chapter 18 includes general principles, production, and detectors of X‐ray waves. Chapter 19 introduces general discussion on quantitative analysis methods and procedures which is the principal subject of XRF spectroscopy. Chapter 20 deals with the crucial aspects concerning the operation and optimization of electronics for X‐ray detection and fluorescence spectrometry.

Part V (Application to Biological Samples) consists of eleven (11) chapters which include the different biological applications of XRF spectrometry. Chapter 21 highlights the theoretical basics of the EDXRF followed by some relevant case studies such as elemental profiling for ionomic studies and food authenticity studies. Chapter 22 deals the application of XRF including TXRF to milk and dairy products. Chapter 23 includes the literature review on the elemental concentration analysis of medical plants using XRF technique. Chapter 24 deals with the application of XRF in animal and human cell biology. Chapter 25 covers a variety of biomedical applications using XRF spectrometry. Chapter 26 describes the usefulness of XRF technique to analyze uranium (U) in blood extracted from wounds. Chapter 27 highlights the use of XRF for the analysis of human hair. Chapter 28 discusses the potential utility of XRF methods to analyze different kinds of biological samples such as calcified/dental tissues, gallbladder and kidney stones, hair, nails, blood, urine, and clinical samples. Chapter 29 describes the principles of using WDXRF for the chemical analysis of plant samples. Chapter 30 covers the use and application of XRF in medicinal biology. Chapter 31 describes in details the use of XRF in pharmacology.

Part VI (Special Topics and Comparison with Other Methods) consists of eight (08) chapters. This section includes some chapter based on special topics and comparisons of XRF with other techniques. Chapter 32 describes the XRF technique and state‐of‐the‐art related techniques specifically as they regard the study of teeth, tartar, and oral tissues. Chapter 33 describes in details the principles, theory and applications of WDXRF spectrometry. Chapter 34 describes the chemometric processing of XRF data which is one of the most important steps in XRF spectrometry. Chapter 35 briefly describes the applications of X‐ray crystallography in medicinal biology. Chapter 36 describes the historical fundamentals of X‐ray instruments and present trends in the field of biological science. Chapter 37, the application and development of XRF spectrometry is discussed for biological objects in Mongolia. Chapter 38 highlights the developments and use of XRF techniques to study arsenic in biological samples in Japan. Chapter 39 is the most important chapter which describes about the current trends and future prospects of XRF technique.

Four chapters (17, 18, 25, and 30) are at very basic level, which will be useful for biologists to understand XRF and its biological applications. The priority is understandable by trading‐off the accuracy or precise expression in these four chapters.

We would like to take this opportunity to express our gratitude to all of the authors for their excellent contributions in this book. We hope the readers will enjoy this book “X‐Ray Fluorescence in Biological Sciences: Principles, Instrumentation and Applications” and that it contributes to the continued instrumental developments of XRF and biological applications. We also hope that it encourages and inspires the beginners to the field in exploring the multifaceted aspects of XRF.

Finally, the critical evaluations and recommendations by the reviewers for the applicability of the XRF methods to biological samples will make this book a valuable asset for anyone employing or improving upon these techniques.

December, 2021

Dr. Vivek Kumar Singh

Professor Jun Kawai

Dr. Durgesh Kumar Tripathi