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Chemical Reagents and Laboratory Methods, Lab Report Example

Pages: 2

Words: 662

Lab Report

IFIU

This report makes an effort in detecting the proteins and peptides through the use of Biuret reagent under laboratory conditions. The illustrations pertaining to colors obtained, presence of peptide bonds and results were presented in the subsequent sections.

Key words: Proteins, Peptides, Pepsin, Trypsin and Chemical Reagents

Detection of Proteins and Peptides using Chemical Reagents and Laboratory methods.

Abstract: Proteins, the prime biochemical compounds, possess numerous polypeptides folded in a globular or fibrous form, play a pivotal role in facilitating biological function. The single linear amino acid polymer chain appears to bind together through peptide bonds amongst carboxyl and amino groups. This report makes an effort in outlining the detection of proteins and peptides in combination or individual basis in the Pepsin and Trypsin enzymes, through the use of Biuret reagent. Twelve tubes segregated in two rows of six each had labeled with the addition of necessary substances as mentioned. The results obtained indicate the presence of peptides and proteins in tubes tested for Pepsin and Trypsin.

Introduction

Pepsin, the principal protein degrading or proteolytic enzyme in the digestive system, is a digestive protease occurring in crystalline form. Trypsin, a serine protease witnessed in digestive system plays an important role in cleaving the peptide bonds especially at the carboxyl side of lysine or arginine (Chapman, 1996). The laboratory question on the detection of peptides and proteins within the sample solutions containing Pepsin and Trypsin has been assessed through the exploration of its relative composition with the use of appropriate chemical reagents. Trypsin, developed by exocrine pancrease assists in protein digestion in stomach and small intestine through the breakdown of polypeptine in to smaller fragments. By catalyzing and hydrolyzing the peptide bonds in the carbonyl groups, the two enzymes (Trypsin and Pepsin) play an important role in digesting the egg albumin (Shawn, 2002). The obtained color helped in estimating the presence of Proteins (Purple), Peptides (Pink) and its combination (Light lavender or Light Purple) in the given sample besides offering relative significance (Randall and Ann, 2009).

Materials and Methods

Laboratory materials include the use of pipettes, burettes, dropper, flasks, chemical reagents and balance. Two rows of six test tubes had positioned in a test tube rack and, the usage of grease pencil had aided in labeling the tubes accordingly. Varying proportions of Pepsin, Trypsin, distilled water, sodium bicarbonate and hydrochloric acid had added to each tube. Numerous pieces of coagulated egg white had placed and checked for signs of digestion for 15minutes. Subsequent to 90minutes of check, 4 drops of Biuret reagent had added to each tube to test for the presence of polypeptides. The color obtained in each tube may be noted and, results obtained had recorded by detailing each experiment.

Results: Table 1: Represents the obtained results with regard to the treatment of Pepsin and Trypsin with Biuret Reagent.

Tube Contents Color INDICATION
1 5ml Pepsin+5ml Distilled water Light Lavender Proteins and Peptides
2 5ml Pepsin + 5ml Sodium Bicarbonate Light Purple Proteins and Peptides
3 5ml Pepsin + 5ml Hydrochloric Acid Pink Peptides
4 5ml Distilled water Purple Proteins
5 5ml Distilled water + 5ml Na2CO3 Nothing Proteins and Peptides
6 5ml Distilled water + 5ml HCl Light Purple Protein
7 5ml Trypsin + 5ml Distilled water Pink Peptides
8 5ml Trypsin + 5ml Sodium carbonate Pink Peptides
9 5ml Trypsin + 5ml HCl Light Blue Proteins
10 5ml Distilled water Purple Proteins
11 5ml Trypsin + 5ml Na2CO3 Light Purple Proteins and Peptides
12 5ml Distilled water + 5ml HCl Blue Proteins

Discussion

The results outlined in the above sections indicate the fact that the use of Biuret reagent subsequent to 90mins of the procedure had helped in detecting the Peptides and Proteins in the given sample. The color obtained for the combination of Proteins and Peptides illustrate the characteristic reaction with the reagent. The hypothesis of detecting the proteins and peptides using chemical reagents under laboratory conditions appear to be significant and relevant despite minimal complexities.

References

Chapman, J.R. Proteins and Peptide analysis by Mass Spectrometry.  Boston: SAGE Publications, 1996, Print.

Doonan, Shawn. Peptides and Proteins. New York: Heinemann Publishers, 2002, Print.

Mrsny, Randall and Daugherty, Ann. Proteins and Peptides: Pharmacokinetic and Pharmacodynamic and Metabolic Outcomes. United States of America: Informa Health Care, 2009, Print.

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