Laboratory Manual for Practical Biochemistry

Ministry of Healthcare of the Russian Federation
theme 1.1. amino acid structUre and classification. Protein structure
	INTRODUCTION
	SELF-STUDY QUESTIONS
		Practical
			COLOR QUALITATIVE PROTEIN AND AMINO ACID REACTIONS
				Reagents
				Material of the investigation
					NINHYDRIN TEST
				Procedure and observation:
				Principle
					TESTS FOR SULPHUR CONTAINING AMINO ACIDS AND PROTEINS
				Principle
				Procedure & observation
				Practical use of color reactions
theme 1.2. sTRUCTURE, PHYSiCAL AND CHEMICAL PROPERTIES OF PROTEINS
	iNTRODUCTION
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical 1
			Reagents
			Material of investigation
			Principle
			Design of laboratory work
		Practical 2
			PROTEIN DENATURATION
				Reagents
				Material forthe investigation
				Principle
					Chemical denaturation.
				Practical significance
				Design of laboratory work
theme 1.3. protein classification. protein structure and functions in human body. complex proteins
	introduction
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical
			Reagents
				Analysis of phosphoprotein chemical composition
			Material of the investigation
				Analysis of glycoprotein chemical composition
			Material for the investigation
			Procedure and observation
			Design of practical
Principle
Principle
Principle
Procedure
Principle
Procedure
Tests
case studies
theme 2.1. FAT SOLUBLE VITAMINS
	INTRODUCTION
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical 1
			QUAliTATIVE RECTIONS OF FAT SOLUBLE VITAMINS
				RETINOL QUALIITATIVE REACTION
				Principle
				Reagents
				Material for investigation
				Procedure and observation
					CHOLECALCIFEROL QUALITATIVE REACTION
				Principle
				Reagents
				Material for investigation
				Procedure and observation
					TOCOFEROL QUALITATIVE REACTION
				Principle
				Reagents
				Material for investigation
				Procedure and observation
					VITAMIN K QUALITATIVE REACTION
				Principle
				Menadione (synthetic analogue of vitamin K1) in the presence of cysteine in a basic medium turns to lemon yellow color.
				Reagents
				Material for investigation
				Procedure and observation
				Clinical and diagnostic significance
				Design of laboratory work
theme 2.2. WATER SOLUBLE VITAMINS
	INTRODUCTION
	SELF-STUDY QUESTIONS
	topics for report
		Practical
			quaLItative reactions OF water soluble vitamins
				Reagents
				Equipment
				Material for investigation
	riboflavin qualitative reaction
		Material for investigation
			Reducing reaction
		Principle
		Procedure and observation
	nicotinic acid qualitative reaction
		Principle
		Material for investigation
		Procedure and observation
	pyridoxine qualitatuvereaction
		Principle
		Material for investigation
		Procedure and observation
	cobalamin qualitative reaction
		Principle
		Material for investigation
		Procedure and observation
	ascorbic acid qualitative reaction
		Material for investigation
		Principle
		Procedure and observation
		Design of laboratory work
tests
case studies
Theme 3.1. ENZYME STRUCTURE AND properties.
APPLICATION IN MEDICINE
	INTrODUCTION
	the aim of practical is
	SELF-STUDY QUESTIONS
	topic for reports
		Practical 1
			dependence of enzyme activity on temperature
				Investigation of influence of temperature on salivary amylase activity
				Reagents
				Material of investigation
				Principle
				Procedure and observation.
					Preparation of saliva (the student on duty performs the saliva for the whole group).
				Design of laboratory work
		Practical 2
			Investigation of enzyme specificity
				Reagents
				Material for investigation
					Detection of urease absolute specificity
				Principle
				Procedure
					Urease preparation:
				Design of laboratory work
					Detection of salivary amylase specificity
				Principle
				Procedure and observation
					Preparation of saliva (the student on duty performs the saliva for the whole group).
				Design of laboratory work
theme 3.2. ENZYME ACTIVITY REGULATION
	INTRoDUCTION
	THE AIM OF THE PRACTICAL IS
	SELF-STUDY QUESTIONS
		Practical
			DETERMINATION OF AMYLASE ACTIVITY IN SERUM and urine
				Principle
				Reagents
				Material for investigation
				Procedure
				Calculation:
				Normal values
				Clinical and diagnostic significance
					Serum and urine
				Design of laboratory work
Theme 3.3.  enzyme classification and nomenclature (seminar)
	introduction
	the aim of the seminar
	SELF-STUDY QUESTIONS
	topic for reports
tests
case studies
Theme 4.1. main catabolic pathways:  pyruvate oxidative decarboxylation. tricarbonic acid cycle. enzymes of respiratory chain. oxidaTive phosphorylation (seminar)
	introduction
	the aim Of practic is
	SELF-STUDY QUESTIONS
TESTS
Case studies
theme 5.1. externalmetabolism of PROTEINs. PRoteinsDIGESTION AND ABSORPTION
	introduction
	the aim of THE practicAL CLASS is
	SELF-STUDY QUESTIONS
	topics for reports
		Practical 1
			qualitative reaction of free hyDrochloric acid in gastric juice
				Material of investigation
					With Congo red
				Principle
				Reagents
				Procedure and observation
					With methyl orange
				Principle
				Reagents
				Procedure and observation
				Normal values
				Clinical and diagnostic significance
				Design of laboratory work
		Practical 2
			qualitative reaction of lactic acid presence in gastric juice
				Principle
				Reagents
				Material of investigation
				Procedure and observation
				Normal values
				Clinical and diagnostic significance
				Design of laboratory work
		Practical 3
			Detection of blood and hemoglobin in gastric juice by diagnostic strips "hemophan"
				Principle
				Material for investigation
				Procedure and observation
				Normal values
				Clinical and diagnostic significance
				Design of laboratory work
		Practical 4 (in theory)
			tubeless method of acidity detection in gastric juice (test of acidity)
				Principle
				Reagents
				Material for investigation
				Procedure and observation
				Design of laboratory work
theme 5.2. intracellular amino acid metabolism
	introduction
	the aim of the practical CLASS is
	SELF-STUDY QUESTIONS
	topics for reports
		Practical
			determination of aminotransferases activity in serum
				Principle
				Reagents
				Material of investigation
				Procedure and observation
				Calculation
				Normal values
				Clinical and diagnostic significance
				Design of laboratory work
theme 5.3. ammonia metabolism and its disposal
	introduction
	the aim of THE practical CLASS is
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			determination of urea content in serum and urine
				Principle
				Reagents
				Material of investigation
				Procedure and observation
				Calculation
				Normal values
				Clinical and diagnostic significance
					Serum
					Urine
				Design of laboratory work
		Practical 2
			determination of creatinine content in serum and urine
				Principle
				Reagents
				Material of investigation
				Procedure and observation
				Calculation
				Normal values
				Clinical and diagnostic significance
					Serum
					Urine
				Design of laboratory work
theme 5.4. metabolism of some amino acids. their features and disorders (seminar)
	introduction
	the aim of THE practical CLASS is
	SELF-STUDY QUESTIONS
	topics for report
tests
case studies
theme 6.1. STRUCTURE AND metabolism of purine and pyrimidine nucleotides
	introduction
	the aim oF the practical CLASS is
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical
			DETERMINATION OF URIC ACID CONCENTRATION IN SERUM AND URINE
				Principle
				Reagents
				Material of investigation
				Procedure and observation
				Calculation
				Normal values
				Clinical and diagnostic significance
					Serum
					Urine
				Design of laboratory work
tests
CASE STUDIES
theme 7.1. nucleic acids synthesis and
its regulation
	introduction
	the aim of THE practical CLASS is
	SELF-STUDY QUESTIONS
	topics for report
		Practical
			Analysis of chemical composition OF nucleoproteins
				Reagents
				Material for investigation
				Procedure and observation
				Design of laboratory work
theme 7.2. protein biosynthesis and its regulation
	introduction
	the aim of THE practicalCLASS IS
	SELF-STUDY QUESTIONS
	topics for reports
		Practical
			determination of protein concentration in serum
				Biuret test
				Principle
				Material for investigation
				Reagents
				Procedure
				Calculation
				Normal values
				Clinical and diagnostic significance
					Hyperproteinemia
					Hypoproteinemia
				Design of laboratory work
tests
case studies
theme 8.1. Structure and metabolism of carbohydrates. Glycogen metabolism
	Introduction
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical
		Detection of glucose in urine using "Glucophan" diagnostic strips
			Design of work:
theme 8.2. Oxidation of glucose in aerobic conditions. Gluconeogenesis
	Introduction
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical 1
			Principle
			Material of investigation
			Reagents
			Procedure
			Normal values
			Clinical and diagnostic significance
		Practical 2
DETECTION OF LACTIC ACID IN MUSCLE TISSUE BY UFFELMAN'S REACTION
	Principle
The method is based on the complex compound interaction of violet iron phenolate with lactic acid to form yellow-green lactate of iron.
	Material for investigation
	Reagents
	Procedure
Theme 8.3. Aerobic oxidation of glucose. Pentose phosphate pathway
	Introduction
	SELF-STUDY QUESTIONS
	TOPICS FOR REPORTS
		Practical
			Principle
			Material for investigation
			Reagents
			The glucose concentration determination
			Calculation
			Normal values
			Evaluation of the glycemic curve
			There are the following types of glycemic curves:
Tests
case studies
theme 9.1. Structure and external metabolism of lipids
	Introduction
	SELF-STUDY QUESTIONS
	topics for reports
		Practical
			Principle
			Material for investigation
			Reagents
			Procedure
			Design of work:
Theme 9.2. Intracellular metabolism of fatty acids and triacylglycerols
	Introduction
	the aim of the practical CLASS IS:
	SELF-STUDY QUESTIONS
	Topics for report
		Practical
			Principle
			Material for investigation
			Reagents
			Procedure
			Normal values
			Clinical-diagnostic significance
			Design of work:
Theme 9.3. Intracellular metabolism of phospholipids and cholesterol. Transport of lipids in blood
	Introduction
	The aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical
			Principle
			Material for investigation
			Reagents
			Procedure
			Normal values
			Design of practical:
Tests
case studies
theme 10.1. The mechanisms of hormonal signal transduction. Classification of hormones. Hormones of pituitary gland. (seminar)
	introduction
	the aim of THE practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
theme 10.2. Hormones of hypothalamus, pituitary, thyroid, pancreas and parathyroid glands
	introduction
	the aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			Material for investigation
			Reagents
			Procedure
	Biuret test for Proteins
		Procedure
	Reaction for (-amino group detection
		Procedure
			Aromatic amino acids reaction
		The principle
		The principle
		Procedure
		Design of practical:
		Practical 2
			Material for investigation
			Reagents
			Procedure
			Design of practical:
Theme 10.3. Hormones of hypophysis, adrenal and sexual glands
	introduction
	the aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical
			The principle
			Material for investigation:
			Reagents
			Procedure
			Design of practical:
Tests
case studies
	Introduction
	the aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			Thymol test
			The principle
			Reagents
			Material for investigation
			Procedure
			Normal values
			Clinical-diagnostic significance
			Design of practical
		Practical 2 (in theory)
			The principle
			Material for investigation
			Equipment
			Procedure (basic steps)
theme 11.2. Iron exchange. Hemoproteins. The synthesis and breakdown of heme
	introduction
	the aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			DETERMINATION OF THE TOTAL BILIRUBIN AND ITS FRACTIONS CONCENTRATION IN BLOOD SERUM
				The principle
				Material for investigation
				Reagents
				Procedure
				Design of practical
		Practical 2
			The principle
			Material for investigation
			Normal values
			Design of practical
theme 11.3. Inorganic substances of the blood.
Acid-base status
	introduction
	the aim of the practical CLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			The principle
			Material for investigation
			Reagents
			Procedure
			Normal values
				Serum
				Urine
			Design of practical
		Practical 2
			The principle
			The material for investigation
			Reagents
			Procedure
			Normal values
			Clinical-diagnostic significance
				Serum
				Urine
			Design of practical
Tests
case studies
theme 12.1. Water-salt exchange. Normal and pathological components of urine
	introduction
	the aim of the practicalCLASS is:
	SELF-STUDY QUESTIONS
	topics for report
		Practical 1
			Material for investigation
			Equipment
			Procedure
			The principle
			Procedure
			Normal value
			The influencing factors
			Design of practical:
		Practical 2
			Material for investigation
			Equipment
			Procedure
			The principle
				Urine
			The principle
			Normal values
				Urine
			The principle
			Normal values
				Urine
			The principle
			Normal values
				Urine
			The principle
			Normal values
				Urine
			Normal values
				Urine
			Design of practical:
Tests
case studies
	Unit 1. "Structure, features and functions of proteins"
	Unit 2. "Structure, classification and role of vitamins"
	Unit 3."Enzymology"
	Unit 4. "Biological oxidation"
	Unit 5. "Amino acid and protein metabolism"
	Unit 6. "Structure and metabolism of purine and pyrimidine nucleotides"
	Unit 7. "Matrix biosynthesis"
	Unit 8. "Structure and metabolism of carbohydrates"
	Unit 9. "Structure and metabolism of lipids"
	Unit 10. "Hormonal regulation of human metabolism and functions"
	Unit 11. "Biochemistry of blood"
	Unit 12. "Biochemistry of kidneys"
	Unit 1. "Structure, features and functions of proteins"
	Unit 2. " Structure, classification and role of vitamins"
	Unit 3. "Enzymology"
	Unit 4. "Biological oxidation"
	Unit 5. "Amino acid and protein metabolism"
	Unit 6. "Structure and metabolism of purines and pyrimidines"
	Unit 7. "Matrix biosynthesis"
	Unit 8. "Structure and metabolism of carbohydrates"
	Unit 9. "Structure and metabolism of lipids"
	Unit 12. "Biochemistry of kidneys"
		The systematic name of oxidoreductases is formed as follows:
		Example 1
		Example 2
		Example 3
	II Class. Transferase
		Group donor : group acceptor – transferable transferase group
		Example 1
		Example 2
		Example 3
	III class. Hydrolase
		1. Esterases – hydrolysis of ester bonds.
		2. Lipases – hydrolysis of neutral fats.
		3. Phosphatase – hydrolysis of monoesters of phosphoric acid.
		4. Glycosidases – hydrolyses O- and N-glycosidic bonds.
		5. Proteases, peptidases – hydrolysis of proteins and peptides.
		6. Nuclease – hydrolysis of nucleic acids.
		The systematic name of hydrolases is formed:
		Hydrolyzable substrate : hydrolase release group
		Example 1
		Example 2
		Example 3
	IV Class. lyase
		The systematic name of lyases is formed:
		Cleavable substrate : detachable group – lyase
		Example 1
		Example 2
		Example 3
		Example 4
	V CLASS. Isomerase
		Example 1
		Example 2
		Example 3
	VI class. ligase
		The systematic name of the enzyme:
		Example 1
		Example 2
		Example 3
		CHARACTERISTICS OFSOME BLOOD PROTEINS
		Fibrinogen
		Normal values
		Clinical and diagnostic significance
	(1-Globulins
		Acid (1-glycoprotein
		Normal values
		Clinical and diagnostic significance
			(1-Antitrypsin
		Normal values
		Clinical and diagnostic significance
			(1-Antichymotrypsin
		Normal values
		Clinicalanddiagnosticsignificance
	(2-Globulins
		С-reactive protein
		Normal values
		Clinicalanddiagnosticsignificance
			Haptoglobin
		Normal values
		Clinical and diagnostic significance
			(2-Macroglobulin
		Normal values
		Clinical and diagnostic significance
			Ceruloplasmin
		Normal values
		Clinicalanddiagnosticsignificance
	(-globulins
		Transferrin family
		Transferrin
		Normal values
		Clinical and diagnostic significance
			Lactoferrin
		Normal values
		Clinical and diagnostic significance
	Normal values of studied
	biocamical indicators
		Serum
		Urine
		Gastric juice
		Clearance of endogenous creatinine
	ContentS