COLORIMETRIC DETERMINATION OF UREA USING DIACETYL MONOXIME

WITH STRONG ACIDS

BY

SADIYA ABUBAKAR

SHT/KN/MLT/RET/21/046

DEPARTMENT OF MEDICAL LABORATORY SCIENCE, SCHOOL OF HEALTH

TECHNOLOGY KANO

JULY, 2023

I
 DECLARATION

“I hereby declare that this work was carried out by me, under the supervision of MLS Habib
Adamu of the Medical Laboratory Science Department, School of Health Technology Kano. I
should be held responsible for any error of omission or commission in the context of the project.”

_______________________ _________________
Sadiya Abubakar Date
SHT/KN/MLT/RET/21/046

II
 CERTIFICATION
This is to certify that the research work titled “DETERMINATION OF UREA USING
DIACETYL MONOXIME WITH STRONG ACIDS” was carried out by Sadiya Abubakar under
my supervision.

_________________________
MLS Habib Adamu

III
 APPROVAL PAGE
This is to certify that the research project report has been examined and approved for the
award of the professional diploma of “MEDICAL LABORATORY TECHNICIAN (MLT)” by
the department of Medical Laboratory Science, School of Health Technology Kano.

………………………………… .……………………………
Project Supervisor Date
MLS Habib Adamu

…………………………………. ……………………………
Project Coodinator Date
MLS

…………………………………. ……………………………
Head of Department Date
MLS Habib Adamu

IV
 DEDICATION
First, I will like to thank to Almighty Allah for giving me strength, courage and ability to
see the end of our program successfully. This project is dedicated to my parents who have taken
all my responsibilities toward the success of my study, may Almighty Allah reward them
abundantly. Ameen.

V
ACKNOWLEDGEMENT

VI
 TABLE OF CONTENTS

TITLE PAGE I

DECLARATION II

CERTIFICATION III

APPROVAL PAGE IV

DEDICATION V

ACKNOWLEDGEMENT VI

TABLE OF CONTENTS VII

ABSTRACT X

CHAPTER ONE 1

1.0 INTRODUCTION 1

1.1 Background of the study 1

1.2 Aim and objectives 3

1.3 Statement of research problem 3

1.4 Significance of the study 3

1.5 Limitations 4

CHAPTER TWO 5

2.0 LITERATURE REVIEW 5

2.1 Urea production and excretion 5

2.2 Clinical significance of urea determination 6

VII
2.2.1. Increases 6

2.2.2 Non-renal cause 7

2.2.3 Decreases 7

2.3 Diacetyl monoxime method of urea determination 7

CHAPTER THREE 11

3.0 MATERIALS AND METHOD 11

3.1 Sample type and collection 11

3.2 Equipment 12

3.3 Reagents and preparations 12

3.3.1 Diacetyl monoxime 4g 12

3.3.2 Urea acid reagent 13

3.3.3 Urea color reagent 14

3.3.4 Urea stock standard 125mmol/L 14

3.3.5 Urea working standard 10mmol/L 14

3.4 Analytical method 15

3.4.1 Principal of the method 15

3.4.2 Procedure of the method 15

3.4.3 Calculations 16

CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

VIII
IX
 ABSTRACT

Urea is a waste product formed in liver following the breakdown of proteins. It passes into the

blood, is filtered out of the kidneys and excreted in urine. Thus determination of blood urea is the

most widely used screening test for the evaluation of kidney function. Numerous analytical

techniques have been developed for the determination of urea. Both direct and indirect methods to

determine urea exist. The aim of this study was colorimetric determination of urea using diacetyl

monoxime (DAM) with strong acids. Therefore, this protocol uses diacetyl monoxime to directly

determine the concentration of urea in patient’s sample. The protocol provides repeatable results

and stable reagents with good color stability and simple measurement techniques for use with a

colorimeter. Urea is colorimetrically determined with diacetyl monoxime, the reaction between

diacetyl monoxime and urea in the presence of sulfuric acid, phosphoric acid, thiosemicarbazide,

and ferric chloride produces a chromophore with a peak absorbance at 520nm, which is directly

proportional to the concentration of the urea present in the sample. The lack of detectable

interferences makes this protocol suitable for the determination of urea in millimolar levels.