ACCRA TECHNICAL UNIVERSITY

SCIENCE LABORATORY TECHNOLOGY

PRACTICAL BIOCHEMISTRY
CLASS 2B2

DATE: 11TH MAY,,2024

Glucose as a Biomarker in Diabetes Diagnosis or Glucose Test used in Clinical Diagnosis

Abstract:Diabetes is a disease in which your blood glucose levels are too high.
This laboratory experiment is aimed to investigate the use of glucose as a biomarker in the
diagnosis of diabetes. The experiment involved analyzing glucose levels in blood(serum) samples
using a glucose test commonly used in clinical diagnosis. The spectophometer was used to
evaluate and assay the amount of glucose in the serum/plasma. The results after showed a clear
correlation between elevated glucose levels and the presence of diabetes, highlighting the
potential of glucose as a reliable biomarker for diabetes diagnosis. The Blood sugar test measured
the amount of a sugar which is the glucose in a sample of your blood.

Introduction:

Glucose is a vital biomarker in the diagnosis and management of diabetes mellitus, a metabolic
disorder characterized by high blood sugar levels. The glucose test, also known as the blood
glucose test, is a widely used clinical diagnostic tool to measure the levels of glucose in the blood.
This report will discuss the role of glucose as a biomarker in diabetes diagnosis, the different types
of glucose tests used in clinical diagnosis, and their significance in patient care.

Glucose as a Biomarker:

Glucose is a primary biomarker for diabetes diagnosis due to its direct correlation with the
disease's pathophysiology. In a normal physiological state, glucose levels are tightly regulated by
hormones like insulin and glucagon. However, in diabetes, this regulation is disrupted, leading to
elevated blood glucose levels. Measuring glucose levels helps clinicians diagnose and monitor
diabetes, as well as assess the effectiveness of treatment plans.

Types of Glucose Tests:

1. Fasting Blood Glucose (FBG) Test: Measures glucose levels after an overnight fast of at least 8
hours.
2. Postprandial Blood Glucose (PPG) Test: Measures glucose levels after a meal.
3. Random Blood Glucose (RBG) Test: Measures glucose levels at any time of day, regardless of
meals.
4. Oral Glucose Tolerance Test (OGTT): Measures glucose levels after consuming a standardized
glucose solution.
5. Glycated Hemoglobin (HbA1c) Test: Measures average glucose levels over the past 2-3 months.

Clinical Significance:
Glucose tests are essential in clinical diagnosis and patient care:

1. Diagnosis of diabetes and prediabetes

2. Monitoring of blood glucose control in diabetic patients
3. Adjustment of treatment plans, including medication and lifestyle changes
4. Identification of hypoglycemia (low blood sugar) and hyperglycemia (high blood sugar) episodes
5. Assessment of glucose metabolism in various physiological and pathological conditions

In conclusion, glucose is a vital biomarker in diabetes diagnosis, and the glucose test is a crucial
tool in clinical diagnosis and patient care. The different types of glucose tests provide valuable
information about glucose metabolism, enabling healthcare professionals to make informed
decisions about patient management and treatment.

Objective
Using spectrophotometer to assay glucose in serum/plasma

Aim
To understand the significance of glucose test in clinical diagnosis

Materials
Serum samples taken at intervals 0, 1. 2 and 2½ hours,Boiling tubes,1.5 ml cuvettes ( for
spectrophotometer reading),Spectrophotometer,Glucose oxidase reagent,Distilled Water ,Glucose
standard

Procedure
Pipette 1000ul of glucose reagent into 3tubes labeled Test, standard and Blank

Pipette 10ul of serum into the Test tube containing the glucose reagent

Pipette 10ul of Glucose standard into standard tube

Pipette 10ul of distilled water into the blank tube

Boil in a water bath for 15mins

Transfer the samples into cuvette

Examine the samples for it absorbance and concentration of the test with spectrophotometer

Results

Discussion
In this experiment, we determined the concentration of glucose in a serum sample using a glucose
reagent test, which is a widely accepted and reliable method for glucose determination (Trinder,
1969). The standard curve was prepared using a glucose standard solution with a known
concentration of 98.02 mg/dL (Sigma-Aldrich, Inc., glucose standard solution, Product Insert),
which served as a reference point for comparison. The average absorbance values for the standard
(98.02 mg/dL), test, and blank samples were 0.097, 0.180, and 0.031, respectively.

The results show that the test sample has a significantly higher absorbance value than the
standard (0.097) and blank samples, indicating a higher concentration of glucose, as expected
based on the principles of spectroscopy (Bergmeyer and Bernt, 1974). The average absorbance
value of the test sample (0.180) is close to the average absorbance value of the standard sample
(0.097), suggesting that the concentration of glucose in the test sample is close to 98.02 mg/dL,
which is the standard concentration used for comparison. This indicates that the test sample has a
glucose concentration within the normal range for healthy individuals (American Diabetes
Association, 2022).

The low absorbance value of the blank sample (0.031) indicates minimal interference from the
reagents and instruments, suggesting a high specificity of the test, which is essential for accurate
glucose determination (Guyton andHall, 2016). The small standard deviation of the absorbance
values for each sample indicates high precision and accuracy of the results, as expected based on
the principles of analytical chemistry (Taylor and Totterman, 2017). This suggests that the
experiment was conducted with a high degree of precision and accuracy, and the results are
reliable and trustworthy.

Based on these findings, we can conclude that the glucose reagent test is a reliable and accurate
method for determining glucose concentration in serum samples, as widely used in clinical practice
(Fuchs and Melmed, 2019). The results suggest that the test sample has a glucose concentration
close to 98.02 mg/dL, which is within the normal range for healthy individuals, according to
established clinical guidelines (American Diabetes Association, 2022). This indicates that the
individual from whom the serum sample was obtained is likely to be healthy and free from any
glucose-related disorders.

Furthermore, the results of this experiment demonstrate the importance of using a standard curve
in spectroscopic analysis, as it allows for the accurate determination of glucose concentration in
serum samples (Bergmeyer and Bernt, 1974). The use of a glucose standard solution with a known
concentration of 98.02 mg/dL (Sigma-Aldrich, Inc., glucose standard solution, Product Insert) as a
reference point for comparison ensures that the results are accurate and reliable.

In addition, this experiment highlights the importance of quality control and quality assurance in
laboratory testing, as small variations in absorbance values can significantly affect the accuracy of
the results (Taylor and Totterman, 2017). The use of a blank sample and a standard sample with a
known concentration of glucose ensures that the experiment was conducted with a high degree of
precision and accuracy, and the results are reliable and trustworthy.

Conclusion :
The consistency in test sample readings (0.180, 0.181, 0.180) demonstrates the reliability of the
experimental procedure. These readings, when compared to those of the glucose standard (0.093,
0.095, 0.098) and tehe blank (0.031, 0.031, 0.031), reveal a significant increase in glucose levels in
the test samples. This consistency suggests hyperglycemia, a hallmark of diabetes, reinforcing
glucose's potential as a diagnostic biomarker (Hill et al…,2016).
Furthermore, the calculated averages for the test (0.180), standard (0.097), and blank (0.031)
samples confirm the validity of the experimental approach. The deviation between the test and
blank sample averages validates that the observed glucose levels are not merely background noise
but indeed reflect elevated glucose concentrations in the test samples (Rampersad, 2012).
Additionally, the known concentration of the glucose standard (98.02 mg/L) provides a reference
point for quantifying glucose levels in the test samples. By comparing the test sample readings to
the established concentration of the standard, a quantitative assessment of glucose levels can be
made, facilitating precise diagnosis of diabetes (American Diabetes Association, 2020).

References:
Hill, N. R., Oliver, N. S., Choudhary, P., and Levy, J. C. (2016). Complications of diabetes.
Endocrinology and diabetes, 23(1), 9-13.
Rampersad, S. N. (2012). Multiple applications of alamar blue as an indicator of metabolic function
and cellular health in cell viability bioassays. Sensors, 12(9), 12347-12360.
American Diabetes Association. (2020). Standards of medical care in diabetes—2020 abridged for
primary care providers. Clinical Diabetes, 38(1), 10-38.
Zheng, Y…,Ley, S..,H., & Hu, F. B. (2018). Global aetiology and epidemiology of type 2 diabetes
mellitus and its complications. Nature Reviews Endocrinology, 14(2), 88-98.