1. CLINICAL
BIOCHEMISTRY LABORATORY
PRESENTED By:
1.Reshma Machamasi
2.Sarmila Timilsina
3.Tisha Shakya
4.Jharana Thapa
5.Karjit Dhami
PRESENTED TO:
Tapeshwar Yadav
(Lecturer)
BMLT, DNHE,
M.Sc. Medical Biochemistry
Department of Laboratory
Medicine, Nobel College of
Medical Sciences,
Kathmandu, Nepal

2.  contest
Contents:
1. Introduction.
2. Objectives.
3. Scope.
4. Basic Equipment Used In Biochemistry lab.
5. Collection And Precaution of Blood.
6. General Tests.
7. Other Tests or Special Tests.
8. Safety Rules In Laboratory.

3. Introduction
Biochemistry is the study of the structure and
function of biological molecules such as proteins,
nucleic acids, carbohydrates and lipids.
Biochemistry is the study of the chemistry of
living things. This includes organic molecules and
their chemical reactions.
 Biochemistry deals with body substance like
enzymes, carbohydrates, amino acids, fats,
proteins, hormones, DNA, RNA, pigments etc.

4. Objectives of biochemistry
The major objective of biochemistry is the complete understanding of
all chemical processes associated with living cells at the molecular
level. Some of the objectives can be listed as follows:
1. Isolation, structural elucidation and the determination of mode of
action of biomolecules.
2. Identification of disease mechanisms.
3. Study of in born errors of metabolism.
4. Study of oncogenes in cancer cells.
5. The relationship of biochemistry with the genetics, physiology,
immunology, pharmacology, toxicology etc.
Biochemistry is related to almost all the life sciences and without
biochemistry background and knowledge, a through understanding of
health and well-being is not possible.

5. Scope:
Biochemists work along with chemists, physicists,
healthcare professionals , engineers and so on.
They can also work for various organisatons including
hospitals , universities , education , drug discovery as well
many more professionals.
They usually involved in research related works.
They can be Biotechnologist , Research Scientist ,
Pharmacologist laboratory technician , Lecturer in an
educational institution etc.

6. Basic Equipments
Used In Biochemistry
Lab

7. 1.Centrifuge:
• It is a laboratory device
that is used for
separation of serum or
plasma from blood.
• Separation of sediment
in urine etc.

9.  A dry bath is
laboratory
equipment that is
used to heat
samples.
 Water bath is used
to incubate sample
in water at a
constant
temperature over a
long period of
time.

10. The device
that measure
the intensity of
concentration
of the colour .

11. It is used to
measure sodium
(Na+),potassium
(K+),chloride
(Cl-),Lithium (Li +)
and Calcium
ion(Ca++).

12.  It perform all
routine tests and
most of special type
of tests.
 Test results are
display or print out.

13. It is used to
transport a measured
volumne of liquid
(reagents or
specimen).

14. Collection and
Precaution of
blood

15.  Mainly there are three types of blood collection
Methods.
1.Capillary puncture method:
2.Arterial puncture method:
3. Vein puncture method:
 For futher testing we need sample mainly blood are
use.For Biochemistry lab we may required large amount
of blood,hence vein puncture should be perform while
collecting blood.

16. Gel tube
EDTA tube
Plane tube

17. 1.Data collection of patient must be done carefully.
2.Container and syringe should be dry. Otherwise, it causes
haemolysis.
3.The blood in the anticoagulant should mixed carefully.
4.Blood collection should not attempts more than three times.
5.The blood should not be force through needle.
6. Plastic tube must not used for calcium test as it adsorbs
calcium.
7. Rapid shaking of tube with blood may cause haemolysis.

18. General Tests Performed
In Biochemistry Lab

19.  Absence or insufficient production of
insulin causes diabetes.
 There are two main types of diabetes:
 Type 1 or Insulin dependent diabetes.
 Type 2 or Non- insulin dependent diabetes.

20. Type 2 or non-insulin dependent diabetes mellitus :
 Type 2 diabetes mellitus,which is much more common occurs when the
body cannot produce enough insulin or the insulin is not working efficiently
enough.
I t was formerly known as non-insulin dependent or adult – onset diabetes
due to its occurrence mainly in people over 40.
# Gestational diabetes mellitus occurs when pregnant women have high
blood sugar levels due to hormones produced in pregnancy.
# A diagnosis of gestational diabetes means a higher risk of developing type
2 diabetes in the future.
Type 1 or insulin dependent diabetes mellitus:
 Type 1 diabetes mellitus occurs when the body cannot produce insulin
which is needed to control blood sugar level.
 These type of diabetes occurs in children hence also known as juvenile
diabetes.

21. Significance:
a) Hyperglycaemia (increased level of glucose ) are seen
in hyperthyroidism , hyperpituitarism , surgical
removal of pancrease , haemorrhage etc.
b) Hypoglycaemia (decreased level of glucose) are seen
in hypothroidism , hypopituitarism , severe exercise ,
starvation etc.
Normal value:
Fasting: 60 -110mg/dl.
Post Prandial: 70- 140 mg/dl.
Random: 70-140 mg/dl.

23.  The Renal function test or kidney
function test refers to a few
commonly used tests to screen for
renal function.
 Analysis of blood and urine samples
can be essential for the evaluation of
kidney (renal) function.
 It includes :
 Urea
 Creatinine
 Uric acid
 Sodium
 Potassium

24.  Urea is the end product of protein metabolism
which is synthesized in liver.
 Increased level of urea are in renal disease,
urinary obstruction,shock burn .
 Decrease level of urea are found in liver failure.
 Normal value: 15-40 mg/dl.

25.  Creatinine is a waste product fromed in muscle
by creatine metabolism.
 It is synthesized in kidney,liver and pancrease.
 Increased condition are in renal
disease,starvation,hyperthyroidism etc.
 Normal value: In male= 0.7 - 1-.4 mg/dl
In female = 0.4 – 1.2 mg/dl

26.  Uric acid is formed in body from the end product of
the metabolism of purines (Adenine and Guanine)
present in RNA and DNA .
 It is a metabolic waste product excreted by kidney .
 Increased level of uric acid (hyperuricaemia) are in
gout ,Renal failure,starvation and so on.
 Decreased level of uric acid (hypouricaemia) are in
liver cirrhosis etc.
 Normal value: In male= 3.5 – 7.0 mg/dl
In female= 3.5 – 6.0 mg/dl

27.  It is a extracellular fluid .
 It maintain heart beat,osmotic pressure and regulates body
acid-base balance.
 Increased level of sodium (hypernatremia) are loss of
water from body , dehydration , diabetes mellitus etc.
 Decreased level of sodium (hyponatremia) are occur due
to diarrhoea , vomitting ,edema, chronic renal disease.
 Normal value: 135-145 mEq/l.

28.  It is a intra cellular fluid .
 It plays important role in tranmission of nerve impulse
and also in cardiac muscle activities.
 Increased level of potassium (hyperkalemia) in renal
failure ,depression ,Anuria etc.
 Decreased level of potassium (hypokalemia) seen in
cardio megaly ,musclar weakness etc.
 Normal valve: 3.5 - 5.0 mEq/litre.

30.  Liver function tests measure
various chemicals in the blood
made by the liver.
 An abnormal result may
indicate a problem with our
liver and may help to identify
the cause.
 Further tests may be needed to
clarify the cause of liver
problem .

31.  Bilirubin
 SGOT(Serum glutamic oxaloacetic transaminase)
 SGPT(Serum glutamate pyruvic transaminase)
 ALP (Alkaline phosphatase )
 Protein
 Albumin
 GGT(Gamma-glutamyl transferase)

32.  It is an orange or yellow pigment formed in the liver
by the break down of haemoglobin and excreted in
bile.
 There are two types of bilirubin.
I. Unconjugated or Indirect bilirubin
II. Congugated or Direct bilirubin.
 Increased in jaundice and haemolysis.
 Normal value:
Total bilirubin=Up to 1 mg%
Direct bilirubin=Up to 0.3 mg%
Indirect bilirubin=Up to 0.7 mg%

33.  It is an enzyme that is normally present in liver
and heart.
 Increased level of SGOT seen in myocardial
infection, liver disease etc.
 Decreased level of SGOT seen in renal dialysis,
vitamin B6 deficiency.
 Normal value: Less than 40 IU/L.

34.  It is an enzyme that is normally present in liver
and heart cells. SGPT is released into blood.
 Increased level of SGPT seen in liver disease
such as cirrosis ,carcinoma, obstructive
jaundice.
 Decreased level of SGPT seen in renal
dialysis.
 Normal value: Less than 40 IU/L.

35.  It is an enzyme present in all tissues of the
body
 It is produced by osteoblast of bone .
 It is localized in cell membrane and with
transport mechanism in liver, kidney and
intestine.
 Increased level of ALP in bone disease and
obstructive jaundice.
 Normal value: 108-306 IU/L.

36.  The amount of proteins found in the fluid
portion of our blood.
 Increased level of total protein in dehydration ,
chronic infection etc.
 Decreased level of total protein in liver
disease,kidney disease,malnutrition etc.
 Normal value: 5.5 - 7.8 gm/dl

37.  Albumin is synthesized in liver and is present in
plasma.
 Albumin transports hormones, fatty acids, and
other compounds, buffers pH and
maintains osmotic pressure among other
functions.
 Increase level of serum albumin seen in
dehydration, multiple myeloma etc
 Decrease level of serum albumin seen in
malnitrition, cirrhosis of liver, nephrptic
syndrome etc

38.  It is a enzyme that transfers gamma-glutamyl functional
groups.
 It is present in the cell membrane of many
tissues,including
kidneys,pancrease,liver,spleen,heart,brain etc.
 GGT is elevated by large quantities of alcohol ingestion.
 Increased level of GGT are in disese of liver , pancrease
etc.
 Normal value: 10-30 IU/L.

39.  A pattern of lipids in the blood.
 Serum lipid profile is measured for
cardiovascular risk prediction and has now
become almost a routine test.
 Fasting blood for 9-12 hours (water only) is
required.
 A lipid profile usually includes the levels of
:
 Total cholesterol.
 High-density lipoprotein (HDL).
 Triglycerides.

40.  It is a type of fat found in our blood.
 They are used to give energy to our body. If we
have extras, they are stored in different places
in case they are needed later.
 Increased level of triglycerides are in risk of
heart disease .
 Normal value: 10-190mg/dl.
Lipemic serum

41.  It is a fat like substance that is found in all
body cells.
 Increased level in obesity, hypothyroidism,
diabetes mellitus,cardiovascular disease.
 Decreased in level Haemolytic jaundice,
intestinal obstruction etc.
 Normal value:150-260mg/dl.

42.  It is good or healthy cholesterol because it
transport cholestrol from tissue to liver.
 Lower levels of HDL cholesterol indicate
increased risk of coronary heart disease.
 Higher levels indicates negative risk factor of
coronary heart disease.
 Normal value: 30 – 60 mg/dl.

43.  LDL is a lipoprotein (a combination of fat and
protein) found in the blood.
 It is called "bad" cholesterol because it picks up
cholesterol from the blood and takes it to
the cells.
 A high LDL level is related to a higher risk of
heart and blood vessel disease.
 Normal value: Upto 150 mg/dl.

45.  It is a endocrine and exocrine functions.
 Pancreatic function tests are useful for
diagonsis of pancreatic disorders.
 Glucagon is produced by α-cells of pancreas
which increase glucose level.
 Insulin is produced by β- cell of pancreas which
decreases glucose level.
 Pancreatic function tests includes:
 Amylase
 Lipase

46.  It is a hydrolytic enzyme which hydrolyses
starch into maltose.
 It presents in saliva and pancreatic juice where
it is secreated by parotid gland.
 Increased level in acute pancreatitis , renal
failure, Appendicitis.
 Normal value:150-340 U/L.

47.  It is a enzyme that catalyses the break down
of fats of fatty acids.
 Lipase is produced in the pancreas, mouth,
and stomach.
 Highamounts of lipase may be found in
the blood when the pancreas is damaged or
when the tube leading from the pancreas
(pancreatic duct) to the beginning of the small
intestine is blocked.
 Normal value: 0 – 160 U/l

48.  It is a collective term
for blood tests used to
check the function of
the thyroid.
 A TFT panel typically
includes thyroid
hormones such as
 Stimulating
hormone (TSH,thyrotropin
)
 Thyroxine (T4)

49. I. T3 (Triiodothronine):
 A free or total triiodothronine test is used to assess
thyroid function.
 It affects almost every physiological process in the
body including growth and development,body
temperature,heart beat.
 Normal value: 1.42 – 4.2 pg/ml.
II.T4 (Thyroxine):
 Free thyroxine (free T4) tests are used to help evaluate thyroid
function and diagnose thyroid diseases,
including hyperthyroidism and hypothyroidism , usually after
discovering that the thyroid stimulating hormone (TSH) level is
abnormal.
 Normal value: 0.65 – 1.97 ng/dl.

50.  A thyroid-stimulating hormone (TSH) blood
test is used to check for thyroid gland
problems.
 The cause of an hypothyroidism, TSH levels
can help determine whether hypothyroidism is
due to a damaged thyroid gland or some other
cause (such as a problem with the pituitary
gland or the hypothalamus).
 Normal value: 0.4 – 5.5 µIU/ml.

51. Free T4 Free T3 Interpretation
High Normal Normal Mild (subclinical)
hypothyroidism.
High Low Normal or
low
Hypothyroidism.
Low Normal normal Mild (subclinical)
hyperthyroidism.
Low High or
normal
High or
normal
Hyperthyroidism.
Low Low or
normal
Low or
normal
Non-thyroidal illness; pituitary
(secondary) hypothyroidism.
Normal High high Thyroid hormone resistance
syndrome (a mutation in the
thyroid hormone receptor
decreases thyroid hormone
function)
TSH

52.  The diagnostic tests in cardiology are methods of
identifying heart conditions associated with healthy vs.
unhealthy, pathologic heart function.
 It includes the tests below:
i. CPK.
ii. CPK MB.
iii. LDH.
iv. GOT/AST.
v. Troponine.
vi. Myoglobin.

53.  Creatine phosphokinase (CPK) is an enzyme in the
body.
 It is found mainly in the heart, brain, and skeletal
muscle.
 This article discusses the test to measure the amount of
CPK in the blood.
2.CPK- MB(isoenzyme of CPK found in heart):
 The CPK-MB test is a cardiac marker used to
assist diagnoses of an acute myocardial infarction.
 It measures the blood level of CK-MB, the bound
combination of two variants (isoenzymes CKM
and CKB) of the enzyme phosphocreatine kinase.

54. 3.LDH(Lactate Dehydrogenase):
 Lactate dehydrogenase (LDH) is a protein that helps
produce energy in the body.
 An LDH test measure the amount of LDH in the
blood.
 It is most often measured to check for tissue damage.
 It is in many body tissues,especially the
heart,liver,kidney,muscles,brain,blood cells and lungs.
4.Troponine:
Troponin is a complex of three regulatory
proteins(troponin C, troponin I, and troponin T ) that
is integral to muscle contraction in skeletal
muscle and cardiac muscle, but not smooth muscle.

55.  Myoglobin (symbol Mb or MB) is an iron- and oxygen-
binding protein found in the muscle tissue of
vertebrates in general and in almost all mammals.
 It is related to hemoglobin , which is the iron- and
oxygen-binding protein in blood, specifically in the red
blood cells .
 In humans, myoglobin is only found in the
bloodstream after muscle injury.

57. Other tests or special tests done in
Biochemistry Lab

58.  It is a major mineral element of body.
 It is present in bones and teeth also present in
body fluids in small concentrations.
 It also involved in blood coagulation.
 Important role in muscle contraction .
 Based source of calcium in milk and their product
 Increased condition is Hypercalcaemia.
 Decreased condition in serum hypocalcaemia.
 Normal value: 8.5-11.0 mg/dl .

59.  Phosphorus is an essential structural component of cell
membranes and nucleic acids.
 Phosphorus is found in most food sources and is a component of
many commonly used food additives.
 Phosphorus is an essential mineral that is required by every cell in
the body for normal function.
 Hyperphosphatemia is observed in fasting then in post
prandial,renal failure,hypervitamininosis,cardiovascular disease
etc.
 Hypophosphatemia is observed in rickets,hypoparathyroidism
etc.
 Normal value: In adult= 2.5 – 5.0 mg/dl.
In children= 4.0 – 7.0 mg/dl.

60.  HbA1c is a form of hemoglobin that is bound to
glucose.
 The blood test for HbA1c level is routinely
performed in people with type 1 and type 2
diabetes mellitus.
 Blood HbA1c levels are reflective of how
well diabetes is controlled.
 The normal range for level for HbA1c is less than
6%.
 HbA1c is also known as glycosylated, or glycated
hemoglobin.

61.  A urine microalbumin test is a test to detect very small
levels of a blood protein (albumin) in urine.
 A microalbumin test is used to detect early signs of
kidney damage in people who are at risk of developing
kidney disease.
 Microalbumin tests are recommended for people with
an increased risk of kidney disease, such as those with
type 1 diabetes, type 2 diabetes or high blood
pressure.
 Normal value: Below 20 mg/dl.

62.  Urine protein test is used to detect protein in the urine
(proteinuria).
 24 hrs urine protein or a random protein to creatinine
ratio may be used to monitor a person with known
kidney disease or damage.
 A urine protein to creatinine ratio may be ordered on
a random urine sample if there is evidence of
significant and persistent protein in the urine.
 Normal value: In male= less than 0.11.
In female= less than 0.16.

63.  The 24-hour urine protein test checks the function of
the kidneys and helps detect disease.
 Urine samples are collected in one or more containers
over a period of 24 hours.
 When higher than normal amounts of protein are in the
urine, it’s called proteinuria. This is often a sign of
kidney damage and disease.
 Normal value: less than 150 mg/day.

64.  Fluid is a liquid that is made in the cavity to lubricate
the surface of the tissue that lines and covers most of
the organs in the cavities.
 It included:
1) Synovial fluid: present in joints like knee joint,hip
joint,elbow joint,shoulder joint etc.
2) Pleural fluid: it is a fluid of pleural surface(around the
lung).
3) Pericardial fluid: fluid present in pericardial
sac(around the heart).
4) Peritoneal fluid: it is a straw coloured clear liquid that
is made in the abdominal cavity.

65.  ADA is a protein that is produced by cells throughout the body
and is associated with the activation of lymphocytes , a type of
white blood cell that plays a role in the immune response to
infections.
 An ADA test may be ordered when a person has an accumulation
of fluid in the chest cavity(pleural fluid) and has signs or
symptoms that suggest TB.
 ADA activity is increased in various conditions such as liver
disease , tuberculosis , typhoid.
 The average of serum ADA in TB and non-TB patients are
20.88U/L and 10.69 U/L respectively.

66. • Cerebrospinal fluid (CSF) analysis is a way of
looking for conditions that affect the brain and
spine.
• It is a series of laboratory tests performed on
a sample of CSF.
• CSF usually contains a small amount
of protein and glucose and may have a few
white blood cells.

67.  A tumor marker is a substance present in or produced by a tumor
or by the tumor’s host in response to the tumor’s.
 Hormones as tumor markers:
a. ACTH
b. ADH
c. Calcitonin
d. HCG etc
 Carbohydrates as tumor marker:
1. CA 15-3
2. CA 125
3. CA 19-9 etc.

68. Name of test Normal test Significance(high)
1.RFT
# Urea 15-40 mg/dl Renal failure, Dehydration, shocks, severe
diarrhea, severe burns, etc.
#Creatinine 0.4-1.4 mg/dl Renal failure, congestive heart failure,
shock,
Obstruction in urinary tract, etc.
#sodium 135 to 145 mEq/L Thirst, fatigue, coma, rapid heart beat,
weakness, etc.
#potassium 3.5 to 5.5 mEq/L Weakness, tiredness, palpitation, etc.
2.LFT
#Bilirubin T.bili= upto 1mg%
D.bili= upto
0.4mg%
Jaundice, haemolysis, etc.
#Total protein 6.3-8.4 gm% Dehydration, multiple myeloma, etc.

69. Name of test Normal value Significance
#ALP 108-306IU/L Pregnancy, hepatic disease, osteoblast, etc.
3.LIPID PROFILE
#Triglyceride Upto 150 mg/dl Diabetes mellitus, liver disease, nephrotic
syndrome, etc.
#Total Cholesterol 150-250 mg/dl Diabetes mellitus, obstructive jaundice,
nephrosis, etc.
#HDL/Good
Cholesterol
30-60 mg/dl Diabetes mellitus, obstructive jaundice,
nephrosis, etc.
#LDL/Bad
Cholesterol
Upto 150 mg/dl Heart and blood vessel disease.
4.TFT
# Free T3 1.42 – 4.2 pg/ml.
#Free T4 0.65 – 1.97 ng/dl.
#TSH 0.4 – 5.5 µIU/ml.

70. Rules for working in a biochemistry laboratory
There are two major concerns to be considered when working in a
biochemistry laboratory. First is safety and second is efficiency in the
laboratory work. There are general rules that we students are advised to
follow:
1. Keep the benches and shelves clean and well-organized.
2. Avoid contaminating the chemicals, use only clean glassware, label
glassware in use.
3. Plan your experiments before starting to carry them out.
4. Pay attention to others in the laboratory.

71. Safety rules in the laboratory
A. Safety rules in general
1. Do not work alone in the laboratory.
2. Unauthorized experiments are not allowed.
3. Eating, drinking and smoking in the laboratory are strictly
prohibited.
4. Become familiar with the location and the use of standard
safety features in the laboratory. The laboratory is equipped
with fire extinguishers, eye washes, safety showers and first-
aid kits. Any question regarding the use of these facilities
should be addressed to your instructor.
5. Special care for eye protection is required. Safety glasses must
be used when certain procedures are being carried out.

72. B. Special safety rules
1. While heating a solution one should make sure not to overheat it.The mouth
of the glassware containing the solution to be heated should never be
pointed toward anyone.
2. Handling of strong acids and bases requires special attention. When diluting
concentrated acids, the acid should be poured into the water and never the
opposite.
3. The pipettes should never be filled with solutions of toxic substances,
biological fluids, strong acids and bases by mouth suction. Use either
automatic pipettes or pipette pumps .
4.Volatile liquids and solids that are toxic or irritating should be handled
under fume hoods.
5. While handling flammable liquids such as ether, alcohols, benzene, naked
flame (burners, matches) must not be in use. The above liquids must not be
stored near radiating heat sources, such as the laboratory oven.
6. Before using electrical appliances, make sure they are grounded.
7. Before leaving the laboratory, electrical equipment should be turned off,
and gas burners extinguished. No tap water should be left running.

73. Thank you !!!!!!