Assignment – Biochemistry

Q 1:- Renal function tests (RFT) are a group of tests that may be performed
together to evaluate kidney (renal) function. The tests measure levels of
various substances, including several minerals, electrolytes, proteins, and
glucose (sugar), in the blood to determine the current health of the kidneys.
Indications for the assessment of renal function are varied and range from
acute emergency to chronic settings.

1. Primarily, renal function tests are performed to identify the renal

disease to determine appropriate patient management and prevent
further deterioration of renal function.
2. Further indications in patients in whom the renal disease has been
identified are to stage level or type of renal disease and to monitor the
progression of renal disease to ensure that optimal management occurs
and to monitor response to interventions [4].

Components of RFT

Clinically, the most practical tests to assess renal function is to get an

estimate of the glomerular filtration rate (GFR) and to check for proteinuria
(albuminuria). [4]

Glomerular Function- Clearance test

 Within the kidneys are about a million tiny blood filtering units called
nephrons. In each nephron, blood is continually filtered through a cluster of
looping blood vessels, called a glomerulus, which allows the passage of water
and small molecules but retains blood cells, proteins such as albumin, and
larger molecules.

The best test to assess glomerular function is Glomerular filtration rate

(GFR), which is the rate in milliliters per minute at which substances in
plasma are filtered through the glomerulus; in other words, the clearance of
a substance from the blood. The normal GFR for an adult male is 90 to 120
mL per minute.

Testing for GFR can be a complicated and lengthy procedure, which is why
doctors use a formula to estimate GFR or eGFR. The standard way to
estimate GFR is with a simple blood test that measures creatinine levels.
Creatinine is a waste product from the digestion of dietary protein and the
normal breakdown of muscle tissue. Aside from chronic kidney disease,
creatinine levels can be affected by other factors, including diet, muscle
mass, malnutrition, and other chronic illnesses.[5]

According to the Kidney Disease Improving Global Outcomes (KDIGO), The

stages of chronic kidney disease (CKD):

 Stage 1 GFR greater than 90 ml/min/1.73 m²

 Stage 2 GFR-between 60 to 89 ml/min/1.73 m²
 Stage 3a GFR 45 to 59 ml/min/1.73 m²
 Stage 3b GFR 30 to 44 ml/min/1.73 m²
 Stage 4 GFR of 15 to 29 ml/min/1.73 m²
 Stage 5-GFR less than 15 ml/min/1.73 m² (end-stage renal disease)
1. Q 2:- any of a class of nitrogenous organic compounds which have large molecules
composed of one or more long chains of amino acids and are an essential part of all
living organisms, especially as structural components of body tissues such as
muscle, hair, etc., and as enzymes and antibodies.
"a protein found in wheat"
o proteins collectively, especially as a dietary component.
"a diet high in protein"
A protein is a naturally occurring, extremely complex substance that consists of
amino acid residues joined by peptide bonds. Proteins are present in all living
organisms and include many essential biological compounds such as enzymes,
hormones, and antibodies.
Protein has many roles in your body. It helps repair and build your body's tissues,
allows metabolic reactions to take place and coordinates bodily functions. In addition
to providing your body with a structural framework, proteins also maintain proper pH
and fluid balance.
 Proteins are large, complex molecules that play many critical roles in the body. They
do most of the work in cells and are required for the structure, function, and
regulation of the body's tissues and organs.
5 Roles of Protein
 Building Tissues and Muscles. Protein is necessary in building and repairing body
tissues. ...
 Hormone Production. Hormones are chemicals produced by glands in one part of the
body that help coordinate activities and communicate with other areas. ...
 Enzymes. ...
 Immune Function. ...
 Energy.
Classification of Proteins
 Simple proteins: They are composed of only amino acid residue. On hydrolysis, these
proteins yield only constituent amino acids. ...
 Conjugated proteins: They are combined with non-protein moiety. Eg. ...
 Derived proteins: They are derivatives or degraded products of simple and
conjugated proteins.
Protein classification based on shape. On the basis of their shape, proteins may be
divided into two classes: fibrous and globular.

Q 3:- Flame photometry is based on the measurement of intensity of the light emitted
when a metal is introduced into a flame. The wavelength of the colour tells us what
the element if, and the colour's intensity tells us how much of the element is present.
The principle of flame photometer is based on the measurement of the emitted light
intensity when a metal is introduced into the flame. The wavelength of the colour
gives information about the element and the colour of the flame gives information
about the amount of the element present in the sample.
It is divided into three types which are absorption, emission, and luminescence
spectroscopy.
A photoelectric flame photometer is a device used in inorganic chemical analysis to
determine the concentration of certain metal ions, among them sodium, potassium,
lithium, and calcium. Group 1 and Group 2 metals are quite sensitive to Flame
Photometry due to their low excitation energies.
These instruments are fairly simple, consisting of four basic components: a flame or
“burner,” a nebulizer (or aspirator) and mixing chamber, color filters, and a photo
detector. Flame photometers are also very cost effective and easy to use.
In 1873, Champion, Pellet, and Grenier developed an instrument that analysed the
content of sodium in plant ash samples to a within 5%, which would be the
“invention” of the flame photometer as we see it today.
 Q 4:- There are four steps involved in obtaining a good quality specimen for
testing: (1) preparation of the patient, (2) collection of the specimen, (3) processing
the specimen, and (4) storing and/or transporting the specimen.
Processing a specimen may include mixing the specimen to ensure that all the
components are evenly distributed throughout the sample or spinning the specimen
in a centrifuge to separate the serum/plasma layer from the red cells.
Most whole blood donations are spun in centrifuges to separate it into transfusable
components: red cells, platelets, and plasma. Plasma may be processed into
components such as cryoprecipitate, which helps control the risk of bleeding by
helping blood to clot. A specimen processor receives and processes specimens such
as blood and urine samples in a laboratory, captures data and handles client and
healthcare practitioner inquiries. Also known as laboratory specimen processor or
Specimen Accessioner. Completely free trial, no card required.
Whole blood can be stored at 4–8°C for up to 24 hours before the serum is separated,
but it must not be frozen. Whole blood should be allowed to clot and then centrifuged
at 1000 × gravitational units (g) for 10 minutes to separate the serum.
Whole blood samples should not remain at room temperature longer than 8 hours. If
assays are not completed within 8 hours, samples should be stored at +2°C to +8°C
no longer than 7 days. If assays are not completed within 7 days, or the sample is to
be stored beyond 7 days, samples should be frozen at -15°C to -20°CAdditives such
as a clot accelerator, anticoagulant, or heparin might be used, depending on the
testing need. Each additive affects blood sample testing and storage in different
ways. For example, hematology procedures often require the blood to remain in the
tube until the anticoagulants stabilize.
. For several decades the standard blood preservative solution consisted of citrate,
dextrose, and, later, phosphate (ACD and CPD). In 1978 a new solution containing
adenine (CPDA-1) was introduced to permit extension of red cell shelf life from 21 to
35 days.

Q 5:- Metabolism is the process by which your body converts what you eat and drink
into energy. During this complex process, calories in food and beverages are
combined with oxygen to release the energy your body needs to function.
Catabolism: The Breakdown
 Stage 1: Glycolysis for glucose, β-oxidation for fatty acids, or amino acid catabolism.
 Stage 2: Citric Acid Cycle (or Kreb cycle)
 Stage 3: Electron Transport Chain and ATP synthesis.
Metabolism refers to all the chemical processes going on continuously inside your
body that allow life and normal functioning (maintaining normal functioning in the
body is called homeostasis). These processes include those that break down
nutrients from our food, and those that build and repair our body.
 The three main purposes of metabolism are: the conversion of the energy in food to
energy available to run cellular processes; the conversion of food to building blocks
for proteins, lipids, nucleic acids, and some carbohydrates; and the elimination of
metabolic wastes.
The processes of making and breaking down glucose molecules are both examples
of metabolic pathways. A metabolic pathway is a series of connected chemical
reactions that feed one another.
The thyroid regulates your metabolism. The two main thyroid hormones are T3 and
T4. Thyroid disorders are common, and they include goiters, hyperthyroidism, and
hypothyroidism.

Q 6:- Most commonly used tests for detection of reducing sugars are Fehling's Test,
Benedict's Test and Barfoed's Test. a) Fehling's Test: Fehling's Solution (deep blue
colored) is used to determine the presence of reducing sugars and aldehydes.
Perform this test with fructose, glucose, maltose and sucrose.
Qualitative examinations measure the presence or absence of a substance, or
evaluate cellular characteristics such as morphology. Semi-quantitative examinations
provide an estimate of how much of the measured substance is present. Qualitative
and semi-quantitative testing must be monitored by quality control processes.
Objective: To characterize carbohydrates present in an unknown solution on the
basis of various chemical assays.
The unspecific Molisch's test for carbohydrates is one of the examples of some tests
which are based on the formation of furfural or furfural derivatives in presence of
concentrated acids. Specific complex formation is sometimes used as specific test
for carbohydrates. Formation of phenylhydrazone is one such example.

Q 7:- Here are some of the different types of glass instruments used in laboratories:
 Bulb and graduated pipettes. These are used to transport specific amounts of fluids
from one place to another.
 Burettes. ...
 Beakers. ...
 Volumetric flasks. ...
 Condensers. ...
 Retorts. ...
 Funnels. ...
 Petri dishes.
Common types of glassware include beakers, flasks, pipettes, and test tubes. Each of
these containers has its own unique form and purpose. Glassware is ideal for lab
settings because glass will not react with chemicals so test results will not be altered,
 glass is transparent for easy monitoring, and glass is heat resistant when using high
temperatures. Pipettes are used to transport a certain amount of fluid from one
location to another.
Examples of glassware used for measurements include:
 Graduated cylinders are cylindrical containers used for volumetric measurements.
 Volumetric flasks are for measuring a specific volume of fluid.
 Burettes are used to disperse precise amounts of liquid reagents.
 From tumblers to champagne flutes, glassware is used to serve water, cocktails,
beer, liquor, wine, coffee, tea and other beverages. With a wide variety of sizes and
shapes, each one has a purpose. Florence: Also known as Boiling Flask is lab
equipment that is ideal for operations that involve heating. ...
 Erlenmeyer Flask: Erlenmeyer Flask is commonly known as conical flasks too. ...
 Sidearm flask: The sidearm flask also goes by the name Buchner.
Firstly, and perhaps a little obviously, glass is transparent which therefore makes
monitoring experiments easier. Glass is also heat resistant and capable of
withstanding thermal shock. It is largely inert and won't react with the substances
used in an experiment.

Q 8:- Vitamin Sources

 Water soluble:
 B-1: ham, soymilk, watermelon, acorn squash.
 B-2: milk, yogurt, cheese, whole and enriched grains and cereals.
 B-3: meat, poultry, fish, fortified and whole grains, mushrooms, potatoes.
 B-5: chicken, whole grains, broccoli, avocados, mushrooms.
In contrast, water-soluble vitamins dissolve in water and are not stored by the body.
Since they are eliminated in urine, we require a continuous daily supply in our diet.
The water-soluble vitamins include the vitamin B-complex group and vitamin C. What
so antioxidants do for the body? Protect body cells and the immune system from
damage that can be done. Why do people need to consume water-soluble vitamins
everyday? Because these vitamins remain in your body for only a short time, you
need them everyday.
There are nine water-soluble vitamins: the B vitamins -- folate, thiamine, riboflavin,
niacin, pantothenic acid, biotin, vitamin B6, and vitamin B12 -- and vitamin C.
Deficiency of any of these water-soluble vitamins results in a clinical syndrome that
may result in severe morbidity and mortality. While the recommended daily amount of
vitamin B-12 for adults is 2.4 micrograms, higher doses have been found to be safe.
Your body absorbs only as much as it needs, and any excess passes through your
urine. High doses of vitamin B-12, such as those used to treat a deficiency, might
cause: Headache.
Serious deficiency may lead to disorders known as beriberi and Wernicke-Korsakoff
syndrome. These disorders are associated with a range of symptoms, including
 anorexia, weight loss, impaired neural function, mental problems, muscle weakness
and heart enlargement. Nutrient deficiencies or diseases can be the result of poor
nutritional intake, chronic health conditions, acute health conditions, medications,
altered nutrient metabolism, or a combination of these factors, and can impact the
levels of both macronutrients and micronutrients in the body.

Q 9:-
a) Hygroscopic refers to a matter's ability to adsorb and absorb water from the
surrounding environment. Some common examples of hygroscopic substances
include: Sodium chloride. Zinc chloride. Calcium chloride. A hygroscopic material
(literally "water seeking") is one that readily absorbs water (usually from the
atmosphere). In most cases, the water can be removed from the material by heating
(sometimes under vacuum or under a flow of dry gas such as nitrogen). Hygroscopic
substances include cellulose fibers (such as cotton and paper), sugar, caramel,
honey, glycerol, ethanol, wood, methanol, sulfuric acid, many fertilizer chemicals,
many salts (like calcium chloride, bases like sodium hydroxide etc.), and a wide
variety of other substances.
b) Minerals are those elements on the earth and in foods that our bodies need to
develop and function normally. Those essential for health include calcium,
phosphorus, potassium, sodium, chloride, magnesium, iron, zinc, iodine, chromium,
copper, fluoride, molybdenum, manganese, and selenium.
The major classes of minerals are given below:
 silicates.
 sulfides.
 carbonates.
 oxides.
 halides.
 sulfates.
 phosphates.
 native elements.
The 5 Most Important Minerals
1. Iron. Yes, I'm starting with the most obvious. ...
2. Calcium. We all know calcium is necessary for strong bones, and it's especially
important for kids. ...
3. Magnesium. Magnesium is crucial for both bone health and energy. ...
4. Zinc. Zinc has a few key jobs. ...
5. Potassium.
Minerals are those elements on the earth and in foods that our bodies need to
develop and function normally. Those essential for health include calcium,
 phosphorus, potassium, sodium, chloride, magnesium, iron, zinc, iodine, chromium,
copper, fluoride, molybdenum, manganese, and selenium.
c) Quality assurance consists of plans, policies, and procedures which together
provide an administrative structure for a laboratory's efforts to achieve quality goals.
Quality assurance has three components: Assessment and monitoring. Development
of the program. Quality improvement (quality control).
Laboratory Quality Assurance (QA) encompasses a range of activities that enable
laboratories to achieve and maintain high levels of accuracy and proficiency despite
changes in test methods and the volume of specimens testedQuality Assurance (QA)
for an analytical laboratory is an essential tool to ensure good comparability of data.
In order to achieve this purpose, processes are mon- itored and performance
problems are systematically solved.
d) Titrimetric analysis is a method of analysis in which a solution of the substance
being determined is treated with a solution of a suitable reagent of exactly known
concentration. The reagent is added to the substance until the amount added is
equivalent to the amount of substance to be determined.. The basic principle of the
titration is the following: A solution – a so called titrant or standard solution – is
added to sample to be analyzed. The titrant contains a known concentration of a
chemical which reacts with the substance to be determined. The titrant is added by
means of a burette.
Types of Titration
 Acid-base Titrations.
 Redox Titrations.
 Precipitation Titrations.
 Complexometric Titrations.
e) Alkaline phosphatase (ALP) is a protein found in all body tissues. Tissues with
higher amounts of ALP include the liver, bile ducts, and bone. A blood test can be
done to measure the level of ALP. A related test is the ALP isoenzyme test.
Abnormally high ALP values may be the result of:
 Bacterial infections.
 Cholangiocarcinoma (bile duct cancer)
 Cholelithiasis (gallstones)
 Biliary obstruction or dyskinesia (lack of motility)
 Bone cancers, including osteosarcoma and chondrosarcoma.
 Bone conditions, including osteomalacia and osteoporosis.

Q 10:- A radioimmunoassay (RIA) is an immunoassay that uses radiolabeled

molecules in a stepwise formation of immune complexes. A RIA is a very sensitive in
vitro assay technique used to measure concentrations of substances, usually
measuring antigen concentrations (for example, hormone levels in blood) by use of
antibodies.
Radioimmunoassay technique (RIA) is a very sensitive in vitro technique used to
measure the concentration of antigens (eg, hormone levels in the blood) through the
use of antibodies directed against these antigens.
The basic principle of radioimmunoassay is competitive binding, where a radioactive
antigen ("tracer") competes with a non-radioactive antigen for a fixed number of
antibody or receptor binding sites.
The advantages of RIA are its relative simplicity and the high sensitivity provided by
the use of radioactive compounds. However, there are several disadvantages as well:
high specific activity-radiolabeled hormones and a scintillation counter are required,
and they may not be easily available.
Radioimmunoassay can be used to measure or determine very small quantity (e.g.
nanogram, picogram) of substances such as antigen, antibodies, hormones, and
drugs. Hence, RIA is used for detection of drugs in forensics, narcotics, and others.
The beta subunit HCG radioimmunoassay is the most sensitive test available. It uses
antisera against the isolated polypeptide chains of HCG to obtain maximum
specificity. Other pituitary hormones do not interfere with this assay.
Radioimmunoassay (RIA) is an in vitro assay that measures the presence of an
antigen with very high sensitivity. Basically any biological substance for which a
specific antibody exists can be measured, even in minute concentrations.

Q11:- Com- puters are used for tutorial-style presentations, simulation of biochemical
reactions, experimental data acquisition and analysis, and for information
management. The history of biochemistry can be said to have started with the ancient
Greeks who were interested in the composition and processes of life, although
biochemistry as a specific scientific discipline has its beginning around the early
19th century. Computer has become very important nowadays because it is very
much accurate, fast and can accomplish many tasks easily. Otherwise to complete
those tasks manually much more time is required. It can do very big calculations in
just a fraction of a second. Moreover it can store huge amount of data in it.

Q12:- The principle of a potentiometer is that the potential dropped across a segment
of a wire of uniform cross-section carrying a constant current is directly proportional
to its length. The potentiometer is a simple device used to measure the electrical
potentials (or compare the e.m.f of a cell). Potentiometer Working Principle
The potentiometer consists of L which is a long resistive wire and a battery of
known EMF V whose voltage is known as driver cell voltage. Assume a primary
circuit arrangement by connecting the two ends of L to the battery terminals. One end
of the primary circuit is connected to the cell whose EMF E is to be measured and the
other end is connected to galvanometer G. This circuit is assumed to be a secondary
circuit.
 The working principle depends on the potential across any portion of the wire which
is directly proportional to the length of the wire that has a uniform cross-sectional
area and current flow is constant. Following is the derivation of used to explain the
potentiometer working principle:
(Ohm’s law)
Where,
I: current
R: total resistance
V: voltage
Where,
⍴: resistivity
A: cross-sectional area
With ⍴ and A constant, I is constant too for a rheostat.
Where,
x: length of potentiometer wire
E: cell with Lower EMF
K: constant
The galvanometer G has null detection as the potential difference is equal to zero and
there is no flow of current. So, x is the length of the null point. Unknown EMF can be
found by knowing x and K.
Since the EMF has two cells, let L1 be the null point length of the first cell with EMF
E1 and L2 be the null point length of the second cell with EMF E2.
Applications of potentiometer
 The potentiometer is used as a voltage divider in the electronic circuit.
 The potentiometer is used in radio and television (TV) receiver for volume control,
tone control and linearity control.
 The potentiometer is used in medical equipment.
 It is used in wood processing machine.

Q 13:- the action or process of using a centrifuge, typically to separate fluids of
different densities or liquids from solids.
"the cells were harvested by centrifugation"
Some common examples of centrifugation include: The extraction of fat from milk in
order to produce skimmed milk. The removal of water from moist lettuce with the help
of a salad spinner. The Spin-drying of water in washing machines in order to remove
water from the clothing.
A centrifuge works by using the principle of sedimentation: Under the influence of
gravitational force (g-force), substances separate according to their density. Different
types of separation are known, including isopycnic, ultrafiltration, density gradient,
phase separation, and pelleting.
The centrifuge works using the sedimentation principle, where the centripetal
acceleration causes denser substances and particles to move outward in the radial
direction. At the same time, objects that are less dense are displaced and move to the
center.
The principle of centrifugation is that the denser particles are forced to the bottom
and lighter particles stay at the top when we spun rapidly. APPLICATIONS: ☆ Used in
daires to separate cream from milk; butter from cream etc.. ☆ Used in washing
machines to squeeze out the water from the wet clothes.