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Biochemical Techniques

Product Details Table of Contents. Table of Contents Chapter 1: Preparation of solutions Chapter 2: Expression of concentration Chapter 3: Buffers and their preparation Chapter 4: Techniques in biochemical evaluation Chapter 5: Carbohydrate estimations Chapter 6: Estimation of lipids Chapter 7: Qualitative and quantitative estimations of amino acids and proteins Chapter 8: Protein purification techniques Chapter 9: Cell disruption and fractionation Chapter Enzymes in metabolism Chapter Isoenzyme analysis Chapter Chromatographic separations Chapter Methods for nutritional quality evaluation of food materials Chapter Nutritional evaluation of forages Chapter Techniques in molecular biology.

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Related Searches. A laboratory Text book of Biochemistry, Molecular Biology. The standard solution of 1 M concentration contains 1 mole of solute per litre of solution. The gram-formula weight of NaCl is Weigh out this quantity of NaCI and dissolve it in water and make its volume to 1 L in a volumetric flask of 1, mL capacity.

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This will be called 1 M solution of NaCI. If we use only one half mole i. Using 2 moles or g in 1 L of solution makes a 2 M solution. To prepare a litre of 0. Hence, use 3. But this is 3. How may we determine the volume of concentrated hydrochloric acid to be measured out which contain 3. The concentrated acid is generally HCl weighs 1. HCl contains: 0. Thus, the volume of solution needed to provide 3. HCl is made to 1 L with distilled water with earlier mentioned information, it will make 0. Reagent 1. In a simple way 1 g dissolved in. Standard solution of hydrated salts: Crystalline salts containing water of hydration must be given special consideration in making up standard solutions.

However, the formula weight of this hydrated salt is This fact has to be taken into consideration when weighing out moles or gram equivalent of such hydrated salts i. Stock solutions are prepared for the substances that are to be used frequently and are stable at higher concentration for several days and can be used after appropriate dilution just before use.

It is sometimes convenient to weigh out a relatively large amount of the compound and prepare a stock solution of that compound from which small amounts can be withdrawn at convenience and added to solution of other components. The use of a stock solution thus cuts down on the amount of pipetting and at the same time reduces variability between a number of similar incubation mixtures, assay mixture etc. A particular volume of solution containing a desired concentration of a substance has to be prepared by using its stock solution.

When a solution contains the solute in an amount in excess of that which can completely be dissolved at a given temperature and the solute in solution is in equilibrium with the excess of undissolved solute, the solution is said to be saturated. The following formula can be applied to compute the volume of desired concentrations.

The use of above formula could be explained by considering the following example. Ordinary glassware should be thoroughly cleaned with washing soda or any detergent followed by washing with ordinary tap water and rinsing with distilled water. Care should be taken to remove previous markings on the glassware, if any while cleaning. Cleaning of oil flasks, used while estimating ether extract, should be done by slight boiling with dilute alkali NaOH followed by same procedure adopted for cleaning ordinary glassware. Care, however, should be taken not to use any brush for cleaning inside the flask to avoid scratch formations.

Graduated glassware may be cleaned by initially keeping in chromic acid solution dissolve about 60 g potassium dichromate in mL tap water by thorough stirring and boiling, to which around mL of commercial sulphuric acid is added slowly after cooling kept in cylindrical jar for reasonable time followed by washing and cleaning as per ordinary glassware.

Discard chromic solution when it develops green colour. Ordinary glassware can be dried by keeping in hot air oven at low temperatures. Graduated glassware pipettes, burettes, measuring cylinders, volumetric flasks etc. Laboratory floor, working tables and water sinks should be kept neat and clean and it should be well ventilated and provided with an exhaust fan to remove unwanted gases, fumes and smoke.

One should work fully protected in laboratory by wearing white drill aprons and shoes. Store chemicals and glassware in alphabetical order in well protected cupboards. Reagents should be properly labelled with date of preparation before placing them on shelf. Systematic breakage record should be maintained. Always use acid and alkali gloves while handling strong acids and alkalies. While working in Kjeldahl digestion room, use fume protecting face mask to avoid inhalation of highly irritating sulphur dioxide fumes.

Distilled water bottles should be kept tightly corked to avoid absorption of atmospheric gases. Always add acid to water slowly from the sides of the container near the sink. While opening liquor ammonia bottles, especially during summer season, cool it for some time in a freezer to avoid sudden spurt of ammonia gas accumulated in the bottles. Set the balance and check the oscillation of pans before using analytical balances. After use the pan and platform of the balance should be cleaned with a camel hair brush, in case of any spillage of chemical, sample etc.

Proper record of usage of special equipment should be made in log book meant for it showing date, time and condition of the equipment. Fire extinguishers should be provided in each laboratory. Always work in company during odd hours for the fear of fire and electrical accidents. All used filter papers and other materials should be deposited in water baskets. Smoking in laboratory premises should be restricted to avoid catching accidental firing by highly inflammable chemicals. All observations should be recorded at least in duplicate. Never pipette strong acids and alkalies with mouth.

Always use adopter or rubber bulb or bulb pipette. Never blow the solution left at the tip of the pipette and delivery of the reagent drawn into pipette should be uniform giving appropriate time, varying from 10 to 30 s. Mouth should be washed quickly with water or weak solution of washing soda during accidental sucking of acids.

Acid and alkali spillage on working tables, floor and clothes should be thoroughly washed with water after suitable neutralising with either weak alkali in case of acid and weak acid in case of alkali. Use always glass distilled water while analysing minerals. No need to use distilled water while making up the volume of digested sample for protein estimation and during the analysis of crude fibre.

Always use self prepared reagents and indicators. Consider lower meniscus for clear colourless and upper meniscus for coloured solutions while recording observations with the help of measuring glassware. During cooling samples in a desiccator, the lid should be displaced to leave a small space, which can be closed after complete cooling. Do not turn on fans during decarbonization of a sample for ashing. Turn on the exhaust during decarbonization and while handling fuming acids and other chemicals.

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Always keep decarbonized samples in a closed container like desiccator while carrying to muffle, otherwise due to light weight the material in silica basin may be displaced due to external air movement. Chapter 2 Expression of Concentration Standard solution: A standard solution is one that contains a precisely known concentration of solute. This section explains the common ways of expressing the concentration of solutions that are required for conducting different experiments.

The molarity of a solution is the number of moles of the solute dissolved per litre of the solution. A solution which contains 1 mole of the solute in 1 L of the solution is called a molar solution. Therefore, for preparing 0.

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Thus, in case of aqueous solutions, 1 molal solution is obtained by dissolving 1 mole of the solute in 1, mL since Sp. A solution having 1 gEq of the solute per litre of solution is called 1 N solution. Therefore, the normality of solution is defined as the number of gram equivalent of substance present per litre of the solution. In the case of acids, it is the number of gram 2.