Techniques for Biochemical Analysis

By Khursheed Hussain, Sameena Maqbool Lone, Khalid Z. Masoodi and Syed Minu Balkhi

Techniques for Biochemical Analysis provides researchers with a practical guide for investigating a variety of different biomolecules. It includes a range of tried and tested protocols, outlining the principles upon which each technique is founded, as well as providing instructions on equipment setup and use, buffer preparation, reagents required, safety considerations and analysis of findings. Beginning with an introduction to biochemistry and laboratory procedures, the book moves on to specific methods focused on investigation of carbohydrates, proteins, enzymes, plant hormones, minerals, amino acids, and more. The large range of protocols covered in this foundational, how-to reference are interdisciplinary and adaptable to a variety of areas, making this an ideal resource for researchers across various fields, including biochemistry, molecular biology, medical sciences, plant physiology, agriculture, and related subjects.

• Features step-by-step methods for biochemical analysis of a variety of compounds
• Explores methods that are applicable and adaptable across a variety of fields, including biochemistry, molecular biology, and related areas
• Provides detailed instructions on how to prepare buffers, the equipment to be used, and the analysis of a variety of molecules, including carbohydrates, lipids, proteins, and hormones
• Contains interdisciplinary and adaptable methods and techniques

• Language: English
• Publisher: Academic Press
• Release date: Apr 4, 2024
• ISBN: 9780443159152

Khursheed Hussain

Khursheed Hussain, has been serving as Assistant Professor at Sher-e-Kashmir University of Sciences & Technology in India for 13 years. He served as Scientific Officer to Vice Chancellor from 2015-2020. He has published more than 120 research articles, 25 review articles and many books. His contribution in the field of research is in varietal and hybrid development as Principal/Co-breeder of 7 vegetable varieties with other varieties in the pipeline. He has been an advisor of more than 50 PG students and has been invited to give many lectures and trainings.

Book preview

Preface

In the intricate landscape of biochemistry, where exploration meets the complexities of analytical methods, Techniques for Biochemical Analysis" stands as a beacon of guidance and knowledge. This comprehensive volume emerges from a recognized need to bridge the gap between theoretical understanding and practical application in biochemical analysis.

The primary audience for this book encompasses undergraduate students venturing into the realms of biochemistry and advanced scholars pursuing MSc and PhD studies. However, this comprehensive guide is not limited to these groups alone. Researchers, educators, and professionals in the field of biochemistry will find this volume invaluable, offering insights and methodologies that cater to their pursuit of knowledge and innovation.

Within these pages lies a wealth of knowledge meticulously curated to facilitate an in-depth understanding of biochemical analysis. The scope of this volume spans from foundational principles to advanced methodologies, offering a comprehensive treatment of diverse topics.

Each chapter presents a meticulous exploration of analytical methods pertaining to carbohydrates, proteins, enzymes, vitamins, phenolics, pigments, toxins, and much more. The organization of this volume is structured to guide readers seamlessly through fundamental concepts to advanced applications, empowering learners to navigate the complexities of biochemical analysis.

Acknowledging the pivotal role of structure in facilitating comprehension, this volume is thoughtfully organized to provide a clear and systematic approach to each topic. The arrangement of chapters facilitates a progressive understanding, ensuring a logical flow of information throughout the volume.

In the ever-evolving landscape of biochemistry, this volume distinguishes itself by offering a comprehensive yet accessible compendium of analytical methodologies. Its uniqueness lies in its ability to cater to diverse academic levels while encompassing a breadth of topics relevant to the field.

We gratefully acknowledge Prof. Nazir Ahmad Ganai, the honorable vice chancellor, SKUAST-Kashmir; Prof. Haroon R. Naik, the director of research, SKUAST-Kashmir; and Prof. M.A.A. Siddique, the director of education, SKUAST-Kashmir, Dean Faculty of Horticulture, SKUAST-Kashmir, and professor and head of the Division of Vegetable Science, SKUAST-Kashmir, whose support and guidance have been invaluable in shaping this volume. His unwavering commitment to fostering academic excellence has been a driving force behind the creation of this comprehensive manual.

May this volume serve as a cornerstone in the pursuit of knowledge, inspiring curiosity, and nurturing a deeper understanding of analytical methods in biochemical analysis.

Khursheed Hussain

Assistant Professor, Vegetable Science

Chapter 1

Introduction to biochemistry

Abstract

The introduction to biochemistry delineates its broad purview, elucidating its intricate connection with molecular biology while highlighting its pivotal role in understanding biological systems. Emphasizing the historical progression from foundational chemical theories to the evolution of biochemistry, the chapter navigates through the diverse branches such as molecular biology, cell biology, metabolism, genetics, and various interdisciplinary fields. Key concepts encompass metabolic processes, cellular functions, genetic variations, and the fundamental importance of biochemistry in addressing medical and biological challenges.

Keywords

Biochemistry; molecular biology; cell biology; metabolism; genetics; interdisciplinary fields; historical progression

Biochemistry is the scientific discipline that focuses on the study of chemical processes and reactions that occur within living organisms. It investigates the structure, function, and interactions of biomolecules such as proteins, nucleic acids, carbohydrates, and lipids, as well as the metabolic pathways that regulate various cellular processes. These biochemical processes, through the orchestration of biochemical signaling and metabolic energy flow, contribute to the remarkable complexity of life. The field of biochemistry has significantly evolved, particularly in the latter decades of the 20th century, becoming indispensable across various disciplines within the life sciences, including botany, medicine, and genetics. Presently, pure biochemistry is primarily concerned with unraveling how biological molecules drive the intricate processes within living cells, thus providing insight into the comprehensive understanding of organisms as a whole.

Biochemistry is closely related to molecular biology, which is the study of the molecular systems that allow genetic information encoded in DNA to result in biological processes. Depending on how the terms are defined, molecular biology might be considered of as a branch of biochemistry, or biochemistry as a tool for investigating and studying molecular biology (Fig. 1.1).

Figure 1.1 Correlation of biochemistry with molecular biology.

Much of biochemistry is concerned with the structures, activities, and interactions of biological macromolecules such as proteins, nucleic acids, carbohydrates, and lipids, which provide the structure of cells and carry out many of life’s operations. The cell’s chemistry is also influenced by the reactions of smaller molecules and ions. These can be inorganic, such as water and metal ions, or organic, such as the amino acids utilized in protein synthesis. Metabolism refers to the processes through which cells obtain energy from their surroundings through chemical reactions.

Biochemistry discoveries are mostly used in medicine, nutrition, and agriculture. Biochemists study illness causes and treatments in medicine. Nutritionists investigate ways to stay healthy and the impact of nutritional deficits. Biochemists in agriculture study soil and fertilizers to find ways to improve crop production, crop storage, and insect management. Much of biochemistry is concerned with the structures and activities of biological components such as proteins, carbohydrates, lipids, nucleic acids, and other macromolecules, though processes are increasingly being prioritized over individual molecules (Fig. 1.2).

Figure 1.2 List of biological components.

Historical background

However, before chemistry could make an acceptable contribution to health and agriculture, it needed to be liberated from immediate practical needs in order to become a pure science. This occurred between 1650 and 1780, beginning with the work of Robert Boyle and finishing with that of Antoine-Laurent Lavoisier, the founder of modern chemistry. Boyle questioned the foundations of his day’s chemical theory and emphasized that the proper goal of chemistry was to establish the composition of substances.

His contemporary John Mayow noticed a basic connection between an animal’s respiration and the burning, or oxidation, of organic materials in air. Then, when Lavoisier conducted his foundational investigations on chemical oxidation, he quantitatively demonstrated the similarity between chemical oxidation and the respiratory process. Another biological phenomenon that piqued the interest of late-nineteenth-century chemists was photosynthesis. The showing by Joseph Priestley, Jan Ingenhousz, and Jean Senebier that photosynthesis is fundamentally the opposite of respiration was a watershed moment in the evolution of biological theory.

Various branches of biochemistry

The main branches of biochemistry are listed below (Fig. 1.3).

Figure 1.3 Various branches of biochemistry.

Molecular biology

It is also known as the Biochemistry roots. It is concerned with the study of the functions of biological systems. This branch of biology discusses the interactions and production of DNA, proteins, and RNA.

Cell biology

The structure and functions of cells in living creatures are studied in cell biology. It is also known as Cytology. Cell biology is largely concerned with the study of eukaryotic species’ cells and signaling pathways, as opposed to prokaryotes, which will be studied under microbiology.

Metabolism

Metabolism is one of the most important processes that all living organisms go through. It is nothing more than the transformations or series of processes that occur in the human body when food is transformed into energy. The process of digestion is one example of metabolism.

Genetics

Genetics is a field of biochemistry that studies genes, their variations, and hereditary traits in living organisms.

Other branches include

Animal and Plant Biochemistry

Biotechnology

Molecular Chemistry

Genetic Engineering

Endocrinology

Pharmaceuticals

Neurochemistry

Nutrition

Environmental

Photosynthesis

Toxicology, etc.

Importance of biochemistry

Understanding the following concepts requires a solid understanding of biochemistry:

1. The chemical processes that convert diet into substances that are specific to the cells of a given species.

2. The enzyme catalytic functions.

3. Making use of potential energy derived from the oxidation of food consumed for the different energy-demanding processes of the living cell.

4. The characteristics and structure of chemicals that make up the framework of tissues and cells.

5. To tackle fundamental medical and biological challenges.

Chapter 2

Introduction to laboratory procedures

Abstract

The introduction to laboratory procedures outlines the essential guidelines and protocols imperative for safe, efficient, and productive laboratory work. Emphasizing punctuality, cleanliness, and meticulous record-keeping, it emphasizes the significance of adhering to safety protocols, maintaining laboratory equipment, and fostering a collaborative environment. The chapter provides a comprehensive list of dos and don’ts, equipment care practices encompassing cleaning, calibration, repairs, and refurbishments, ensuring the optimal functionality and safety of laboratory equipment.

Keywords

Laboratory procedures; safety protocols; equipment care; dos and don’ts; laboratory guidelines; record-keeping; equipment maintenance

A laboratory is a space for students, researchers, professors, and scientists to conduct experiments, devise new methodologies for the preparation of chemical substances, and discover new techniques for understanding science. A solid understanding of the operating principles of various types of equipment as well as the properties of the chemicals is required to ensure that they are handled appropriately and safely.

Instructions to work in the laboratory

1. Punctuality should be the primary concern and priority.

2. Your attitude toward your employment will be reflected in your results.

3. Keeping discipline in the laboratory.

4. The laboratory environment should be always kept tidy and clean.

5. All glassware, chemicals, and laboratory equipment should be stored in the specified area.

6. The work area should always be clean. Books, backpacks, and other items should never be left on the workbench. Nothing should be left on the bench unless it is necessary for the experiments.

7. Talking and eating should never be permitted in the laboratory while working. Food should never be allowed in the laboratory.

8. A realistic basic record, field book, and pencil or pen are required to document the experiment’s activities/results. When working in a laboratory, one should always wear a laboratory coat, head hat, mask, gloves, and laboratory shoes.

9. Results should be recorded on time at all times. If there is an issue, the laboratory in charge should always be approached and asked to solve it.

10. Every calculation and step in the experiment protocol should be documented in your field book.

11. Work should always be organized in such a way that it is completed within the time frame specified.

12. Be frugal with your reagents and other resources. Reagents should only be used in minimal amounts. The prepared compounds should be stored at the appropriate temperature.

13. Glassware should be handled with extreme caution. Any breakage should be notified to the responsible laboratory in charge.

14. All liquid waste should be disposed off in the sink, and water from the tap should be left running for a few minutes to drain the waste.

15. It should be ensured that no chemical is ever spilled in or on the laboratory equipment. If this is the case, the equipment should be cleaned as soon as possible following usage.

16. When not in use, all power supplies must be turned off.

17. When not in use, fans, lighting, air conditioners, and computer systems should be turned off.

18. Water supply should always be tightly closed after usage.

19. In the event of an injury or burn, professional medical attention should be provided, and a first aid box should always be kept in the