Clinical Biochemistry

By
Asst. Lec. Nagham Saadi Mohammed
M.Sc. Microbiology

naghamsaadi94@yahoo.com
Clinical Biochemistry

 Clinical Biochemistry is the division of laboratory medicine

that deals with the measurement of chemicals (both natural and
unnatural) in blood, urine and other body fluids.

 These test results are useful for detecting health problems,

determining prognosis and guiding the therapy of a patient.
Principles of Spectrophotometry
The amount of light passing through the tube is measured by the photometer.
The photometer delivers a voltage signal to a display device, The signal changes as
the amount of light absorbed by the liquid changes.
 Kidney function Tests

 One of their most important jobs is to filter waste materials from

the blood and expel them from the body as urine.
 The kidneys also help control the levels of water and various
essential minerals in the body. In addition, they’re critical to the
production of:
•red blood cells
•hormones that regulate blood pressure
Symptoms of kidney problems
Symptoms that may indicate a problem with your kidneys include:
o high blood pressure
o blood in the urine
o frequent urges to urinate
o difficulty beginning urination
o painful urination
o swelling of the hands and feet due to a buildup of fluids in the body
Types of kidney function tests
Serum creatinine test
This blood test examines whether creatinine is building up in your blood. The
kidneys usually completely filter creatinine from the blood. A high level of
creatinine suggests a kidney problem.

Type of specimen collection

 Serum or plasma
Requirement and procedure
 Creatinine testing kit ( R1+R2)
R1= Picric acid
R2=Alkaline Reagent
 Spectrophotometer
 Timer
 Water bath/heat block (37 C)
AS 2 -AS 1
X con. of standard = mg/dl
AST2 -AST.1

Normal value in serum

0.6-1.1 mg/dL men
0.5-0.9 mg/dL women
Blood urea nitrogen (BUN)
 The blood urea nitrogen (BUN) test also checks for waste
products in your blood.
 BUN tests measure the amount of nitrogen in the blood. Urea
nitrogen is a breakdown product of protein.
 However, not all elevated BUN tests aredue to kidney damage.
Commonmedications, including large doses of aspirin and some
types of antibiotics, can also increase your BUN. It’s important to
tell your doctor about any medications or supplements that you take
regularly.
Specimen collection
Serum or heparinised plasma
Required Equipment
 Water bath at 37ºC , Timer
 Spectrophotometer at 600 nm.
 Urea testing kit

• for Procedures of any kit , you should follow manufacturer’s instructions.

• Incubated at 37 ºC for 5-10 minutes and read at 600 nm
• Calculations of results

AS
X concentration of standard = mg/dl
AST
 Uric acid
• Uric acid is the final product of purine metabolism , which found in the
nucleic acid .
• Analysis or testing of uric acid levels is used for the diagnosis and
treatment of numerous renal disorders including renal failure and kidney
stones.
• Specimen collection
Serum or plasma
Requirements
• Water bath at 37ºC , Timer
• Spectrophotometer at 520 nm
• Uric acid testing kit
Procedures For kit methods, follow manufacturer’s instructions.
Incubated at 37 ºC for 5-10 minutes and read at 520 nm
Result / Calculations of results

As
X con. of standard = mg/dl
Ast
 Glucose

Glucose is the main source of energy in the body. The levels of this
compound are balanced by digestion and absorption of carbohydrates in the
intestine, its storage and release in the liver and its utilisation in the muscle.
Apart from the screening for and monitoring of diabetes, glucose is measured
in cases of pancreatic, metabolic or endocrinic disorders.

Requirements
- Glucose testing kit
-EDTA , Heparinized serum , plasma ,CSF.
- Spectrophotometer
- Water bath (optional) at 37ºC
Incubated at 37 ºC for 5-10 minutes and read at 500 nm

Result / Calculations of results

X con. of standard = mg/dl

Normal values:
Fasting blood sugar : 70-100 mg/dl
Random Blood sugar : 70-120 mg/dl
HbA1c
 The term HbA1c refers to glycated haemoglobin. It develops when
haemoglobin, a protein within red blood cells that carries oxygen
throughout your body, joins with glucose in the blood, becoming
‘glycated’.
 By measuring glycated haemoglobin (HbA1c), clinicians are able to get
an overall picture of what our average blood sugar levels have been over
a period of weeks/months.
 For people with diabetes this is important as the higher the HbA1c,
the greater the risk of developing diabetes-related complications.

 The amount of glucose that combines with this protein is directly

proportional to the total amount of sugar that is in your system at that
time.

 Because red blood cells in the human body survive for 8-12 weeks
before renewal, measuring glycated haemoglobin (or HbA1c) can be
used to reflect average blood glucose levels over that duration,
providing a useful longer-term gauge of blood glucose control.

 If your blood sugar levels have been high in recent weeks, your
HbA1c will also be greater.
 Lipid profile test

 Cholesterol
 Triglyceride
 HDL : High density lipoprotein
 LDL : low density lipoprotein
 VLDL very low density lipoprotein
 cholesterol
 A cholesterol test can help determine your risk of
the buildup of fatty deposits (plaques) in your arteries
that can lead to narrowed or blocked arteries
throughout
 A cholesterol test is an important tool. High
cholesterol levels often are a significant risk factor for
coronary artery disease.your body (atherosclerosis).
Requirements
• Water bath at 37ºC , Timer
• Spectrophotometer at 500 nm
• cholesterol testing kit
Incubated at 37 ºC for 5-10 minutes and read at 500 nm

Result / Calculations of results

X con. of standard = mg/dl

Normal values:
Less than 200 mg/dl
 Triglyceride
Triglycerides are a type of fat (lipid) found in your blood.
When you eat, your body converts any calories it doesn't need to use right
away into triglycerides. The triglycerides are stored in your fat cells.
Later, hormones release triglycerides for energy between meals.
Incubated at 37 ºC for 5-10 minutes and read at 500 nm

Result / Calculations of results

X con. of standard = mg/dl

Normal values:
Less than 150 mg/dl
• HDL stands for high-density lipoproteins. It is sometimes called the
"good" cholesterol because it carries cholesterol from other parts of your
body back to your liver. Your liver then removes the cholesterol from your
body.
• LDL stands for low-density lipoproteins. It is sometimes called the "bad"
cholesterol because a high LDL level leads to a buildup of cholesterol in
your arteries.
• VLDL stands for very-low-density lipoprotein. Your liver makes VLDL
and releases it into your bloodstream. The VLDL particles mainly carry
triglycerides ,another type of fat, to your tissues. VLDL is similar to
LDL cholesterol but LDL mainly carries cholesterol to your tissues
instead of triglycerides.
 VLDL = Triglyceride /5 mg/dl

LDL= Total cholesterol – HDL –(TG/5)

 What are liver function tests?

Liver function tests, also known as liver chemistries, help determine

the health of your liver by measuring the levels of proteins, liver enzymes,
and bilirubin in your blood.
liver function test is often recommended in the following
situations:
 to check for damage from liver infections, such as hepatitis B and
hepatitis C
 to monitor the side effects of certain medications known to affect the
liver
 if you already have a liver disease, to monitor the disease and how well a
particular treatment is working
 if you’re experiencing the symptoms of a liver disorder
 if you have certain medical conditions such as high triglycerides, diabetes
,high blood pressure, or anemia
 if you drink alcohol heavily
 if you have gallbladder disease
Commonly used tests to check liver abnormalities are tests checking:
• alanine transaminase (ALT)
• aspartate aminotransferase (AST)
• alkaline phosphatase (ALP)
• albumin
• bilirubin
 Alanine transaminase (ALT)

 Alanine transaminase (ALT) is used by your body to metabolize

protein. If the liver is damaged or not functioning properly, ALT can be
released into the blood. This causes ALT levels to increase.
 A higher than normal result on this test can be a sign of liver
damage.
Aspartate aminotransferase (AST)

 Aspartate aminotransferase (AST) is an enzyme found in several

parts of your body, including the heart ,liver, and muscles. Since AST
levels aren’t as specific for liver damage as ALT, it’s usually measured
together with ALT to check for liver problems.
 When the liver is damaged, AST can be released into the
bloodstream. A high result on an AST test might indicate a problem
with the liver or muscles.
 The normal range for AST is typically up to 40 IU/L in adults and
may be higher in infants and young children.
Alkaline phosphatase (ALP)

 Alkaline phosphatase (ALP) is an enzyme found in your bones, bile

ducts, and liver. An ALP test is typically ordered in combination with
several other tests.
 High levels of ALP may indicate liver inflammation, blockage of the
bile ducts ,or a bone disease.
 Children and adolescents may have elevated levels of ALP because
their bones are growing.
 Pregnancy can also raise ALP levels. The normal range for ALP is
typically up to 120 U/L in adults.
 Albumin

 Albumin is the main protein made by your liver. It performs many

important bodily functions.
 An albumin test measures how well your liver is making this particular
protein.
 A low result on this test can indicate that your liver isn’t functioning
properly.
 The normal range for albumin is 3.5–5.0 grams per deciliter (g/dL).
However, low albumin can also be a result of poor nutrition, kidney
disease, infection, and inflammation.
 Bilirubin test

 Bilirubin is a waste product from the breakdown of red blood cells

.It’s ordinarily processed by the liver. It passes through the liver before
being excreted through your stool.
 A damaged liver can’t properly process bilirubin. This leads to an
abnormally high level of bilirubin in the blood. A high result on the
bilirubin test may indicate that the liver isn’t functioning properly.
 The normal range for total bilirubin is typically 0.1–1.2 milligrams
per deciliter (mg/dL). There are certain inherited diseases that raise
bilirubin levels, but the liver function is normal.
 The ALT and AST tests measure enzymes that your liver releases in
response to damage or disease.
 The albumin test measures how well the liver creates albumin, while
the bilirubin test measures how well it disposes of bilirubin.
 ALP can be used to evaluate the bile duct system of the liver.
 Having abnormal results on any of these liver tests typically requires
follow up to determine the cause of the abnormalities. Even mildly
elevated results can be associated with liver disease. However, these
enzymes can also be found in other places besides the liver.
Cobas C111