Techniques for Biochemical Analysis

1st Edition - April 4, 2024
Authors: Khursheed Hussain, Sameena Maqbool Lone, Khalid Z. Masoodi, Syed Minu Balkhi
Language: English
Paperback ISBN:
9 7 8 - 0 - 4 4 3 - 1 5 9 1 4 - 5
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 1 5 9 1 5 - 2

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 pr… Read more

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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.

Cover image
Title page
Table of Contents
Copyright
Preface
Chapter 1. Introduction to biochemistry
Abstract
Historical background
Various branches of biochemistry
Chapter 2. Introduction to laboratory procedures
Abstract
Instructions to work in the laboratory
Dos and don’ts in the laboratory
Laboratory equipment care
Chapter 3. Preparation of solutions and buffers
Abstract
Common definitions
Preparation of solutions
Preparation of buffers
Mechanism of buffering action
Buffering capacity
Applications of buffer in chemistry
Chapter 4. Qualitative analysis of carbohydrates
Abstract
Classification of carbohydrates
Tests for carbohydrates
Chapter 5. Qualitative analysis of proteins
Abstract
Classification of carbohydrates
Functions of proteins
Chemical tests for the estimation proteins
Further reading
Chapter 6. pH (negative logarithm of hydrogen ion concentration)
Abstract
Measurement of pH
pH indicators
Measurement of pH
Chapter 7. Spectrophotometry
Abstract
Rationale
Qualitative photometric assays
Quantitative photometric assays
Standard curve
Measurement of absorbance
Application of spectrophotometry
Chapter 8. Estimation of carbohydrates
Abstract
Sample extraction for estimation of sugars
Estimation of total sugars by anthrone method
Estimation of total sugars by phenol−sulfuric method
Estimation of reducing sugars by Nelson−Somogyi method
Estimation of nonreducing sugars
Estimation of starch by anthrone reagent method
Estimation of glycogen by anthrone reagent method
Reference
Chapter 9. Amino acids and proteins
Abstract
Amino acids
Proteins
Functions of proteins
Nitrogen estimation by micro-Kjeldahl method
Nonprotein nitrogen estimation
Protein nitrogen estimation
Amino-nitrogen estimation
Estimation of amino acids
Chapter 10. Enzyme assays
Abstract
Determination of amylase activity
Determination of invertase (β-fructofuranosidase) activity
Determination of ribonuclease activity
Determination of deoxyribonuclease activity
Determination of β-glucosidase activity
Determination of pyruvate decarboxylase activity
Determination of urease activity
Determination of lipoxygenase (lipoxidase) activity
Determination of phospholipase C activity
Determination of α-galactosidase activity
Determination of nitrate reductase (NR) activity
Determination of nitrite reductase activity
Determination of catalase activity
Determination of peroxidase activity
Determination of lactate dehydrogenase activity
Determination of chitinase activity
Determination of polygalacturonase activity
Determination of pectin methyl esterase activity
Determination of L-phenylalanine ammonia lyase activity
Determination of tyrosine ammonia lyase activity
Determination of polyphenol oxidase or DOPA oxidase activity
Determination of phytase activity
Determination of malate dehydrogenase activity
Determination of indoleacetic acid oxidase activity
Determination of ascorbic acid oxidase activity
References
Chapter 11. Vitamin assays
Abstract
Estimation of vitamin B1 (thiamine)
Estimation of Vitamin B2 (riboflavin)
Estimation of vitamin B3 (niacin)
Estimation of vitamin B6 (pyridoxine)
Estimation of Vitamin A (retinol)
Estimation of Vitamin C (ascorbic acid)
Estimation of vitamins D2 and D3
Estimation of Vitamin E (tocopherol)
Estimation of choline
Chapter 12. Phenolics
Abstract
Estimation of total phenols
Estimation of ortho-dihydric phenols
Estimation of flavanols
Estimation of tannins
Estimation of lignins
Estimation of capsaicin
Chapter 13. Plant pigments
Abstract
Estimation of total carotenoids
Estimation of lycopene
Estimation of chlorophyll
Estimation of curcumin
Estimation of anthocyanin
Estimation of leuco-anthocyanin
Reference
Chapter 14. Plant acids
Abstract
Estimation of oxalic acid
Separation and identification of organic acids using chromatography
Chapter 15. Antinutritional factors in plants
Abstract
Estimation of trypsin inhibitor activity
Estimation of chymotrypsin activity
Estimation of α-amylase inhibitor activity
Estimation of cyanogen’s
Estimation of Gossypol
Estimation of phytic acid
Chapter 16. Toxins
Abstract
Estimation of aflatoxins
Chapter 17. Extraction and identification of polyacetylenes
Abstract
Rationale
Reagents required
Protocol
Analysis and statistics
Chapter 18. Alkaloids
Abstract
Estimation of solanine
Purity assessment of solanine using chromatography
Chapter 19. Plant hormones
Abstract
Estimation of auxins (indole-3-acetic acid)
Estimation of gibberellins
Estimation of cytokinins
Estimation of abscisic acid
Estimation of ethylene (C2H4)
References
Chapter 20. Immunochemical methods
Abstract
Enzyme-linked immunosorbent assay
Chapter 21. Nitrogenase activity
Abstract
Estimation of nitrogenase activity
Chapter 22. Chlorophyll
Abstract
Estimation of chlorophyll
Chapter 23. Proximate analysis
Abstract
Determination of dry matter and moisture content
Determination of ash content
Determination of acid-soluble and -insoluble ash content
Determination of crude fiber
Determination of crude fat
Determination of crude protein
Determination of nitrogen-free extract
Chapter 24. Minerals
Abstract
Macronutrients
Micronutrients
Estimation of sulfur (S)
Estimation of phosphorus
Estimation of calcium
Estimation of magnesium
Estimation of potassium
Estimation of molybdenum
Estimation of copper
Estimation of zinc
Estimation of manganese
Estimation of iron
Estimation of boron
Estimation of cobalt
Estimation of chlorine
Estimation of silica
Estimation of sodium
References
Chapter 25. Antinutritional factors in plants
Abstract
Nutritional studies by in vitro methods
Nutritional studies by in vivo methods
Chapter 26. Antioxidants
Abstract
Estimation of FRAP
Scavenging activity of DPPH radical
Scavenging activity of ABTS radical
Chapter 27. Lipids
Abstract
Estimation of lipids
Index

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.

Affiliations and expertise

Assistant Professor, Division of Vegetable Science. Sher-e-Kashmir University of Sciences& Technology, India

Sameena Maqbool Lone

Sameena Maqbool Lone is a PhD scholar in the Division of Vegetable Science, Sher-e-Kashmir University of Agricultural Sciences and Technology, India. She holds a Master's degree in Horticulture from SKUAST-K, Shalimar. At the Master's level, she worked on "Heterosis and Combining Ability Studies in Cherry Tomato" and obtained competence in morphological and molecular characterization, as well as numerous plant tissue culture technologies. In her Phd programme, she is working on the “Morpho-Molecular Characterization and Nutraceutical Profiling of different carrot genotypes (Daucus carota L.)”. She has outstanding academic and extracurricular achievements, for which she received a merit scholarship from SKUAST-K during her Bachelor's and Master's degree programmes. She has contributed to over 23 academic papers, 6 books, and more than 100 new gene sequences in Genbank by 2022.

Affiliations and expertise

Division of Vegetable Science, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, India

Khalid Z. Masoodi

Dr. Khalid Zaffar Masoodi is a young scientist who works as an Assistant Professor in the Division of Plant Biotechnology at the Sher-e Kashmir University of Agricultural Sciences and Technology in Kashmir (India). He holds a Master's degree in Biotechnology and a doctorate in Plant Biotechnology. Dr. Masoodi is the recipient of numerous awards, including the SERB Early Career Research Award 2016, the SBUR Travel Award in the United States of America in 2012, and the Society of Endocrinology Journal Award in 2018 at Glasgow United Kingdoms. Dr. Masoodi has a distinguished academic record, having qualified for the CSIR-UGC net in Life Sciences, the DBT-BET JRF, and the Graduate Aptitude Test in Engineering and Sciences (GATE) while doing his Masters in Biotechnology. Dr. Masoodi has filed Six patent applications in last 2 years, has over 12 years of teaching and research experience, and has published numerous scientific articles in the fields of cancer research, human disease research, and biotechnology research. His laboratory (Transcriptomics Laboratory, often known as K-Lab, after the Klenow fragment) at SKUAST-K is fully equipped with fluorescence imaging, real-time PCR, cell culture, and Western blotting including other state-of-the-art equipment. His research interests span multiple dimensions, including transcriptomics of plant stress biology (biotic/abiotic) and cancer biology. Apart from that, he is extensively involved in the research of new drugs from medicinal plants. Additionally, he is a co-author of Advanced Molecular Biology and Biotechnology Methods (Academic Press, 2020).

Affiliations and expertise

Assistant Professor/Junior Scientist at the Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, India

Syed Minu Balkhi

Dr. Syed Minu Balkhi holds a PhD in Biochemistry from the University of Kashmir, India. She qualified for the JRF-NET CSIR and obtained a fellowship during her PhD program in 2013. She is currently a Lecturer at Government Boys Higher Secondary School, Humhama, India. Prior to this she was a Post-Doctoral Associate at the Department of Immunology, University of Pittsburgh, USA. She has contributed to various publications.

Affiliations and expertise

Lecturer, Government Boys Higher Secondary School