PREPARATION OF BUFFER SOLUTION: “TRIS BUFFER SOLUTION”

PREPARATION OF BUFFER SOLUTION: “TRIS BUFFER SOLUTION”

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ABSTRACT

Tris buffer solution was prepared in the laboratory by dissolving 4.6g of boric acid, 6.5g of EDTA and 60.5g of trisma base in 1000cm3 of distilled water. Comparative study were done on the commercially produced tris buffer solution, using and laboratory produced tris buffer solution, using pH meter for the p H and by carrying out genotype test for the efficacy of Tris buffer solution prepared in the laboratory was used as an electrolyte. This genotype test reduces the occurrence of sickle cell anemia. This expresses the efficacy rate and pH of laboratory produced Tris buffer solution as compared with that of commercially produced tris buffer solution. The result obtained for the pH of laboratory produced tris buffer solution was found to be 9.0 while that of commercially produced tris buffer solution was found to be 8.9. the result obtained for the efficacy using genotype test showed that tris buffer solution produced in laboratory has more efficacy than that produced commercially, but the difference is not significant.

CHAPTER ONE

1.0            INTRODUCTION

Martin et al (1989) defined buffer as a solution that has the ability to resist change in pH, when small amounts of strong acids or bases are added to it. For example, when 0.01 mole of strong acids or base is added to distilled water, the pH drops to 2 with the acids and rises to 12 with the base. If the same amount of acid or base is added to an acetic acid-sodium acetate buffer, the pH may only change a fraction of a unit.

Buffers are important in many areas of chemistry. When the pH must be controlled during the course of a reaction, the solutions are often buffered. This is often the case in biochemistry when enzymes or proteins are being studied. Our blood is often buffered to a pH of 7.4. Variations of a few tenths of a pH unit can cause illness or death. Acidosis is the condition when pH drops too low. Alkalosis is the condition when pH becomes higher than normal. Acidosis or alkalosis affects the functions of the heart.

In buffer solution two species are required one is capable of reaction with OH- and the other will react with H3O+. The two species must not react with each other. Many buffers are prepared by combing a weak acid and its conjugate (acetic acid and sodium acetate) or a weak base and its conjugate (ammonia and ammonium chloride). In general, the pH range in which a buffer solution is effective is + or – one pH unit on either side of the pka.

Buffering however is the tendency of a solution to resist change in pH following addition of acid or base, according to Martin et al, 1989.

1.1     CONSTITUENT OF A BUFFER

A buffer is a solution made up of a weak acid and its conjugate base, similarly a buffer solution may be made up of a weak and its conjugate acid.

EXAMPLES:

ACID                                                                           CONJUGATE

1.       CH3COOH (acetic acid)                                        CH3COO-

2.       CH3NH                                                                      CH3NH2

                         OH       

3.                                                                                       O-

                             (Phenol)

   

4.      

      HN+                NH

                                                                           N                   NH  

          Base                                                            Conjugate acid

1.       NH2   (ammonia)                                                   NH3+          

2.       CH3- COO- (acetic acid)                                        CH3 COOH

3.       CH2-  COO- (Glycine)                                       CH2 - COOH 

          NH2                                                                                                           NH3+

          Tris buffer solution is a solution that is used in routine analysis of genotype test.  It is used as an electrolyte.  Tris buffers are used by biochemists to control pH in the physiological range (about 7 to 8 pH).

1.2     STATEMENT OF PROBLEM

                The research project tends to produce tris buffer solution that have desired or needed efficacy.  The need for the efficacy of this tris buffer solution is to ensure accurate result when used diagnostically in genotype test.  Genotype test is the test carried out to find out the genotype of individuals as far as sickle cell anaemia is concern.  Sickle cell anaemia is a congenital disease that affects over 30% to 40% of Nigerians.  It is a devastating lifetime disease that has no cure.  However, carrying out genotype test between individuals that are about to get married can prevent it.  In order to avoid getting offspring with sickle cell anaemia, these problems and many others prompted to this research work

1.3     OBJECTIVES OF THE RESEARCH

          The objective/aim of this research work is to produce the tris buffer solution that has required efficacy in the laboratory, also to know the constituents and working pH range of the tris buffer solution. Consequently, to know the diagnostic use of tris buffer solution

1.4     HYPOTHESIS

Ho      -        Tris buffer solution produced in the laboratory has the same

                   efficacy as commercially produced Tris buffer solution.

H1      -        Tris buffer solution produced in the laboratory do not have the

                   same efficacy as commercially produced Tris buffer solution.

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