June 2014 - BIOSCIENCE EDUCATION

Latest Science Discoveries, Reagents Preparations, Standard Laboratory Analysis Procedures and Promoting the importance of Life to the World

Friday 27 June 2014

Preparation of Simulated Intestinal Fluid (SIF)

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Preparation of Simulated Intestinal Fluid (SIF)

Preparation of Simulated Intestinal Fluid (SIF)

Simulated Intestinal Fluid is a synthetic fluid use in analysis of pharmaceuticals in the laboratory. The extent to which the analysed pharmaceutical dissolve and release it actives in the simulated intestinal is a good indicator of how the pharmaceutical will be available for absorption into the system from the villi in the intestine. The Simulated Intestinal Fluid is use to determine bioavailabilty of a particular drug when taken.

Procedure

1.      Weigh 6.8g of monobasic potassium phosphate into a 500ml beaker

2.      Add 250ml of distilled water and mixed to dissolve.

3.      Add 77ml of 0.2N NaOH.

4.      Transfer the solution into 1000ml volumetric flask

5.      Add 300ml of water

6.      Add 1g of Pancreatine to the solution.

7.      Measure the pH (The pH will be in region of 4.5-4.6 at 25oC)

8.      Correct the pH of the solution to pH 6 by adding 0.2N NaOH to increase the pH value to 6.0 (0.2N HCl can also be used if the solution is basic).

9.      Make up the solution to the 1000ml mark.

Monday 23 June 2014

Preparation of 0.5M Acetate buffer

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Preparation of 0.5M Acetate buffer

Preparation of 0.5M Acetate buffer

1.      Weigh 2.99g of sodium acetate trihydrate into a beaker

2.      Transferred into 1000ml volumetric flask with half filled with distilled water

3.      Add 1.66ml of glacial acetic acid

4.      Make up to the mark with distilled water.

5.      The pH of the solution should be measured with a pH meter and should be 4.51  at 37C.

Friday 20 June 2014

Preparation of Simulated Gastric Fluid (SGF)

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Preparation of Simulated Gastric Fluid (SGF)

Preparation of Simulated Gastric Fluid (SGF)

 

Simulated Gastric Fluid (SGF) is a synthetic form of the gastric fluid in the stomach. This fluid will show the effect of the gastric juice in the stomach on a particular drug in the stomach.

Drugs made for dissolution and disintegration in the small intestine must not disintegrate or dissolve in the in the Simulated Gastric Fluid (SGF). On the other hand, drugs designed to act in the stomach will dissolve in this synthetic solution of Simulated Gastric Fluid (SGF).

 

Procedure

To prepare Simulated Gastric Fluid (SGF)

1.      Weigh 2g of NaCl and 3.2g of Pepsin derived from stomach mucosa with an activity of about

2.      800 to 1200 units per milligram

3.      Add 7ml concentrated HCl to dissolve

4.      Transfer into 1000ml volumetric flask

5.      Add deionized/distilled water into 1000ml volumetric flask in drops

6.      Make up to the mark with deionized/distilled water.

7.      Test the pH with a pH meter and the pH should be a pH of 1.199.

Wednesday 18 June 2014

Preparation of 0.2N Phosphate buffer of pH 5.8

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Preparation of 0.2N Phosphate buffer of pH 5.8

Preparation of 0.2N Phosphate buffer of pH 5.8

1.      Weigh 54.44g of potassium dihdro-orthophosphate (KH2PO4) into a beaker

2.      Transfer and washed into a 2000ml volumetric flask with deionized/distilled water

3.      Make up to the mark with deionized/distilled water.

4.      Measure 1250ml of the solution into 5000ml beaker to read the pH.

5.      The pH value should be measured with a pH meter and the pH should be in the region of 4.21

6.      Increase to 5.8 with 0.2M NaOH in drops with continuous stirring.

7.      Transferred the solution into a 5 litre volumetric flask

8.      Make up to the mark with deionized/distilled water (pH 7).

PRAPARATION OF FEHLING’S SOLUTIONS

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PRAPARATION OF FEHLING’S SOLUTIONS

PRAPARATION OF FEHLING’S SOLUTIONS

Fehling solutions are vital reagent for Biochemist and Chemists for qualitative and quantitative analysis of carbohydrates in the laboratory. It was first prepared by Hermann Von Fehling.

The following are the standard procedure for the preparation of Fehling I and II solutions.

Preparation of Fehling I solution

APPARATUS: 500ml Beaker, stirrer, 1000ml volumetric flask

REAGENT: Distilled water, CuSO4 powder

PROCEDURE:

1.      Weigh 69.3g of CuSO4 powder into a 500ml beaker, add 200ml distilled water and stir to dissolve

2.      Transfer to 1000ml volumetric flask, make up to the mark with distilled water and shake to homogenise

 

Preparation of Fehling II solution

REAGENT: Distilled water, NaOH pellets, Potassium sodium tartarate powder

PROCEDURE:

1.      Weigh 100g of NaOH pellets into a 500ml beaker, add 200ml distilled water and stir to dissolve

2.      Weigh 346g of potassium sodium tartarate into another 500ml beaker, add 300ml distilled water and stir to dissolve

3.      Transfer both solutions to 1000ml volumetric flask, make up to the mark with distilled water and shake

NB: The above preparations are for the preparation of 1 litre of the resulting solution.

Thursday 5 June 2014

PREPARATION OF 0.1M AgNO3

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PREPARATION OF 0.1M AgNO3

PREPARATION OF 0.1M AgNO3

AgNO3 also called silvertrioxonitate(iv) is a vital salt in analysis in inorganic chemistry.

Silvertrioxonitrate is a good reagent in qualitative analysis of chloride ions in solution of their salts.

Procedure

Weigh 17g of the AgNO3 powder into a 5oml beaker, dissolve with little amount of distilled water, transfer into 1000ml volumentric flask and make up to the mark with distilled water.

NB: Do not allow the AgNO3 to touch any part of your body and the preparation must be carried out in a dark room.

Monday 2 June 2014

PREPARATION OF 4% BORIC ACID WITH SCREENED METHYL RED

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PREPARATION OF 4% BORIC ACID WITH SCREENED METHYL RED

PREPARATION OF 4% BORIC ACID WITH SCREENED METHYL RED

This is a reagent used in the analysis of protein in the Kjedahl method for collecting the condensed ammonia during distillation.

It is a useful weak acid for analysis of organic substances

Procedure

The procedure to prepare 1000ml 4% Boric acid with screened methyl red is as follows:

APPARATUS: 500ml Beaker, stirrer, 1000ml volumetric flask

REAGENT: Deionized water, Boric acid powder, Bromo cresol green, methyl red

PROCEDURE:

1.      Weigh 40g of Boric acid (B(OH)3) powder into a 500ml beaker, add 200ml deionized water and stir to dissolve

2.      Transfer to 1000ml volumetric flask,

3.      Add 10ml Bromocresol green, 7ml Methyl red, and make up to the mark with deionized water and shake.