ASSIGNMENT

Subject Code PDA103T

Subject Name Medicinal Biochemistry
Programme/Cours
e
PHARM D
Department PHARMACY PRACTICE
Faculty PHARMACY

Name of the Student ANTO AMITH BIJU

Reg. No 16PHPH010005
Semester/Year 1st YEAR/2016
Subject Leader/s MRS. MAMATA.K

i
 M. S. Ramaiah University of Applied Sciences
University House, Gnanagangothri Campus, New BEL Road,
M S R Nagar, Bangalore, Karnataka, INDIA - 560 054.

<Subject Title> ii
 Declaration Sheet
Student Name ANTO AMITH BIJU
Reg. No 16PHPH010005
Programme/Course PHARM D Semester/Year 1st YEAR/2016
Subject Code PDA103T
Subject Title MEDICINAL BIOCHEMISTRY
Subject Date 22/8/16 to 18/1/17
Subject Leader Mrs.MAMATA.K

Declaration

The assignment submitted herewith is a result of my own investigations and that I have
conformed to the guidelines against plagiarism as laid out in the Student Handbook.
All sections of the text and results, which have been obtained from other sources, are
fully referenced. I understand that cheating and plagiarism constitute a breach of
University regulations and will be dealt with accordingly.

Signature of the Student Date

Submission date stamp

(by Examination & Assessment
Section)

Signature of the Subject Leader and date Signature of the Reviewer and date

iii
Faculty of Pharmacy
M.S. Ramaiah University of Applied Sciences
Department Pharmaceutical Chemistry
Programme Pharm D Course PHARM D
Batch 2016 Year I Pharm D
Course Code PDA103T Course Title Medicinal Biochemistry
Course Leader(s) Mamatha K

Assignment
Reg.No. 16PHPH010005 Name of Student ANTO AMITH BIJU

Assignment
Marks
Sections

Marking Scheme Maximum First Second

marks Examiner Examiner

1.1 Presentation skill 2

1.2 Knowledge on concept 3
A 1.3 Appropriate usage of edible oils 3
1.4 Citation and References 2
Part-A Max Marks 10

1.1 Identification of pollutants or pesticides 4

B.1 1.2 Mechanism 4
1.3 Citation and References 2
B.1 Max Marks 10

2.1 Knowledge on laboratory interferences 4

B.2 2.2 False positive 4
2.3 Citation and References 2
B.2 Max Marks 10

Total Assignment Marks 30

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 Course Marks Tabulation

Component-1 (B) First Second

Remarks Remarks
Assignment Examiner Examiner
A
B.1
B.2
Marks (Max 30 )
Marks (out of 15 )

Signature of First Examiner Signature of Second Examiner

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 Scheme of Evaluation

Total
Marks
Allotted
Question Tasks –Steps Allotted
Instructor’s Expected Solution Marks
No. involved for
for the
steps
question
A 1.1 Presentation There are a wide variety of cooking 10
2
oils from plant sources such as olive
skill
oil, palm oil,soybean oil, canola
oil (rapeseed oil),  corn oil, peanut
1.2 Different kinds oil and other vegetable oils, as well as
animal-based oils like butter and lard.
of cooking oils, 4
Deep frying also increases the
their ingredients
formation of advanced glycation end
and health products (AGE’s) on the surface of
foods that, eventually triggers the
benefits.
4 formation of some carcinogens such
1.3 Add a note on
as acrylamides (found in
proper cooking carbohydrate-rich foods such as
French fries), heterocyclic amines and
methods of edible
polycyclic aromatic hydrocarbons
oils emphasizing 2
(chemicals formed when meat is
on appropriate cooked at high temperatures),
aldehyde’s and acrolein, all increases
heating
the risk of cardiovascular deaths.
temperatures
Oils with high smoke points, such as
1.4 Reference corn, soybean, peanut and sesame,
are good for high-heat frying and stir-
frying. Olive, canola and grapeseed
oils have moderately high smoke
points, making them good for sauteing
over medium-high heat.
Oils with low smoke points, such as
flaxseed and walnut, are best saved

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 for use in salad dressings and dips.
B 1.1 Identify any 4 The most important known inhibitors 10

five pesticides, of the ETC are Amytal, Rotenone,

insectides or Antimycin A, CO, Sodium Azide, and

pollutants that Cyanides.

affects biological Amytal, a barbiturate, and Rotenone,

oxidation a plant product used as insecticide

1.2 Mechanism of and pesticide, block the ETC between

4
toxicity NADH dehydrogenase (Complex I)

and CoQ.

Rotenone and MPTP (a neurotoxin),

when administered in vein, cause at

the same time interference with the

functioning of Complex I and a

Parkinson-like disease. These

substances affect primary neurons in

substantia nigra; causing impairment

of Complex I, impairment of

mitochondria metabolism,

accumulation of free radicals, cell

death, release of toxic compounds

and destruction of other cells.

Antymicine A interferes with electron

flow from cytochrome bH in Complex

III (Q-cytochrome c oxidoreductase).  

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 Carbon monoxide (CO) is responsible

for more than 50 % of death by

poisoning worldwide. High levels can

result from incomplete combustion of

fuels: engine and furnace exhausts,

tobacco smoking is important

sources.

Carbon monoxide intoxication causes

impaired oxygen delivery and

utilization at the cellular level. As a

consequence of the binding of CO to

heme molecules, the heart

1.3 References
functioning is impaired leading to

severe hypotension and death.

Cyanide compounds include

hydrogen cyanide gas and the

crystalline solids potassium cyanide

and sodium cyanide. The cyanide ion

halts cellular respiration by inhibiting

2
the enzyme cytochrome c oxidase.

Inhaled cyanide causes coma with

seizures, apnea, and cardiac arrest,

followed by death in seconds.

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 2.1 Liver functions
Liver enzyme levels may be affected
tests
by a variety of factors other than liver

disease, including pregnancy, periods

of rapid bone growth, alcohol

4
consumption, obesity, strenuous

exercise, consumption of particular

foods, trauma, and surgery. 10

Elevated levels of hepatic markers

2.2 False positive
may be seen in these conditions
results
giving false positive results.

Bilirubin may be increased due to

2.3 References
B 4
hemolysis, intra-abdominal bleed,

hematoma other than in hepatitis

AST may be elevated in skeletal

2 muscle injuries, cardiac issues,

hemolysis

ALT may be elevated in Skeletal

muscle, cardiac muscle, renal issues

LDH may be elevated in cardiac

issues, hemolysis

Alkaline phosphatase is also raised in

first trimester, bone, kidney and

intestinal disorders

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<Subject Title> x
<Subject Title> xi
 Contents
____________________________________________________________________________

Declaration Sheet.....................................................................................................................................ii
Contents...................................................................................................................................................iii
List of Tables.............................................................................................................................................v
List of Figures...........................................................................................................................................vi
List of Symbols........................................................................................................................................vii
Question No. 1..........................................................................................................................................8
1.1 Overview:........................................................................................................................................8
1.2 Solution to the question:................................................................................................................8
1.3 Discussions /Suggestions/Views/Recommendations......................................................................8
1.4 Conclusions.....................................................................................................................................8
Question No. 2..........................................................................................................................................9
2.1 Overview:........................................................................................................................................9
2.2 Solution to the question:................................................................................................................9
2.3 Discussions /Suggestions/Views/Recommendations......................................................................9
2.4 Conclusions.....................................................................................................................................9
Question No. 3........................................................................................................................................10
3.1 Overview:......................................................................................................................................10
3.2 Solution to the question:..............................................................................................................10
3.3 Discussions /Suggestions/Views/Recommendations....................................................................10
3.4 Conclusions...................................................................................................................................10
Question No. 4........................................................................................................................................11
4.1 Overview:......................................................................................................................................11
4.2 Solution to the question:..............................................................................................................11
4.3 Discussions /Suggestions/Views/Recommendations....................................................................11
4.4 Conclusions...................................................................................................................................11
Question No. 5........................................................................................................................................12
5.1 Overview:......................................................................................................................................12
5.2 Solution to the question:..............................................................................................................12
5.3 Discussions /Suggestions/Views/Recommendations....................................................................12
5.4 Conclusions...................................................................................................................................12

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PART A

1.1
Name of the Method of Ingredients Smoke point / Uses of the oil Benefits
oil extraction temperature

Saturated fat: 87g Used for Cures Uterine

Coconut oil Dry or wet Polyunsaturated cooking diseases and
process fatty acid: 1.8g 177°C Non- dairy heart diseases
Monounsaturated cream PreventsArthritis
fatty acid :6g NSAID
alternative
Mono Useful for hair Balances
Walnut oil Cold pressing unsaturated fatty 160° and skin care immune system
acid:23g Cuts
Polyunsaturated inflammation
fatty acid:63g Prevents eczema
Saturated acid: 9g

Saturated Useful Prevent diabetes

Canola oil Expelling acid:15g 200° forcooking Improves
process Polyunsaturated meat products reproductive
fatty acid:45g health
Mono Lowers blood
unsaturated fatty cholesterol
acid:11g
Saturated fat: 12g Removes tan Increase
Mustard oil Distillation Polyunsaturated 254° and spots appetite
fat: 21g Act as hair Cardiovascular
Monounsaturated vitalizer benefits
fat: 59g Reduces risk of
cancer
Reduces
Fat: 100 g Protection cholesterol level
Peanut oil Expeller Polyunsaturated: 227°C against forming Reduces from
pressing 32 g wrinkles risk of heart
Saturated: 17g diseases
Monounsaturated Strengthens
: 46 immune system
g
Saturated acid : Prevent arthritis
Sunflower oil Cold process 13g Used in Prevent from
Polyunsaturated 227°C cosmetic colon cancer
fatty acid:36g Formulation Reduces heart
Monounsaturated Frying oil disease risk
fatty acid:46g

Saturated acid : 207° Cooking oil Reduces the

Olive oil Expeller 14g cholesterol level
processing Mono Prevent cancer

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 unsaturated fatty
acid :73g
Poly unsaturated
fatty acid :11g

Ricin oleic Used in Boosts immunity

Castor oil Cold process acid:85% 200° cosmetics Anti-bacterial
Oleic acid: 2 % Beneficial for Reduces arthritis
Linoleic acid: 1 % hair and skin
Linolenic acid:
0.5%
Mono Improves
Almond oil Oil pressing unsaturated fatty Anti-aging circulatory
acid:31g 216° Function as a function
Polyunsaturated carrier oil for To overcome
fatty acid:12g other flavours hormone
Saturated imbalance
acid:3.7g

Grapeseed oil Mono

Cold pressing unsaturated fatty Prevents acne Anti-allergic
acid:17g 204° &stretch marks Cures oedema &
Polyunsaturated Reduces hair varicose veins
fatty acid:71g fall & dandruff
Saturated acid:
12g

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1.2
Deep frying is essentially the most original and one of the crucial oldest approaches of meals
practise global. It includes heat and mass transfer. To decrease the fees, the oils are typically used
again and again for frying. When heated again and again, changes in physical look of the oil will
occur such as accelerated viscosity and darkening in colour, which may alter the fatty acid
composition of the oil. Heating reasons the oil to bear a sequence of chemical reactions like
oxidation, hydrolysis and polymerization. For the period of this approach, many oxidative products
equivalent to hydroperoxide and aldehydes are produced, which can be absorbed into the fried food.
High heat destroys oils (polyunsaturated faster than others) and the by-products of breakdown
don't taste excellent and are very likely carcinogenic. Even though a high temperature oil can be
used at low temperature (sacrificing flavour) it is not advised to use an oil larger than its rated
variety.
Cooking with vegetable oils releases poisonous chemical substances linked to cancer and different
ailments like melanoma, coronary heart disease, dementia and cancer, in step with leading
scientists.
Braising, broiling and grilling, poaching, deep frying etc. are some healthy cooking methods
advised to retain flavour and nutritional value of food without addition of excess amount of oils or
fats.
http://www.skillsyouneed.com/ps/fats-oils.html
http://www.intechopen.com/books/lipid-peroxidation/repeatedly-heated-vegetable-oils-and-lipid-
peroxidation
http://www.seriouseats.com/2014/05/cooking-fats-101-whats-a-smoke-point-and-why-does-it-
matter.html

PART B
INTRODUCTION
The electron transport chain is a set of membrane-embedded proteins and natural and
organicmolecules, most of them organized into four huge complexes labelled I to IV.

15
Because the electrons travel by means of the chain, they go from a higher to a slash vigour degree,
relocating from less electron-hungry to extra electron-hungry molecules. Vigour is launched in
these “downhill” electron transfers, and a couple of the protein complexes use the launched energy
to pump protons from the mitochondrial matrix to the intermembrane house, forming a proton
gradient.
The sequence of reactions within the electron transport chain yields a proton gradient which is
required for the discharge of ATP from ATP synthase. These procedures are coupled. Therefore, if
an inhibitor of the electron transport chain have been present, it will negatively have an impact on
the creation of ATP.
PESTICIDES THAT EFFECT BIOLOGICAL OXIDATION:
 PYRIDABEN(Complex 1 inhibitor) :

Pyridaben is an insecticide and acaricide which is accepted to be used inside the European
and plenty of other countries. It has a highly volatile and low aqueous solubility, based on
its chemical houses, is not predicted to leach to groundwater. It tends not to persist in soils
or water structures. It is fairly toxic to mammals and now not predicted to bioaccumulate. It
is incredibly toxic to maximum aquatic organisms and honey bees but much less so that you
could earthworms and birds.

Pyridaben (PYR), a extensively used acaracide, to be the most amazing compound at

causing oxidative damage and cellular dying, both of which can be attenuated by way of the
antoxidants α-tocopherol and coenzyme Q10. Further, PYR become pretty toxic to midbrain
organotypic slices. Those outcomes spotlight the capability of typically used pesticides to
cause toxicity through similar mechanisms as ROT
Pyridaben inhibited respiratory in bugs in vivo and blocked breathing at complicated I in
numerous mitochondrial arrangements in vitro. Pyridaben additionally blocks complex I and
the induction of ornithine decarboxylase in human breast most cancers cells in vitro an
movement that correlates with antiproliferative and anti most cancers activity.

 MEPRONIL (Complex 2 inhibitor)

Mepronil is a systemic fungicide with shielding and curative actions in a vast range of plants
against basidiomycete fungi through seed treatment, soil and foliar packages. It isn't always
mutagenic in well known exams. It has a completely low toxicity to birds and aquatic
vertebrates and invertebrates. A sequence of substituted benzanilides including Mepronil

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 was shown to inhibit succinic dehydrogenase pastime with large efficiency in isolated
fungal mitochondria. The oxidation of NADH become no longer inhibited.
This end result cautioned that mepronil selectively inhibits the complex II place however
not complexes I, III and IV regions in the electron shipping device. It inhibited the
following enzymes within the remoted mitochondria from R, solani which are responsible
for the oxidation of succinate; succinate-cytochrome c reductase, succinate-coenzyme Qio
reductase, succinate hydrogenase and succinate dehydrogenase. It, however, did not inhibit
succinate dehydrogenase which was solubilized from the mitochondria. It had no
considerable impact on succinate dehydrogenases in mitochondria isolated from different
resources including Pyricuraliaoryzae, Botrytis cinerea, rat liver, mouse liver, etiolated pea
seedling and sweet potato root. Those consequences indicated that mepronil selectively
inhibits succinate dehydrogenase in mitochondria of R. Solani.

 FENAZAQUIN(Complex 1 inhibitor):

Fenazaquin is an ascaricide with broad spectrum action available under the trade names of
Demitan, Totem, and Boramae etc. Fenazaquin causes slight irritation of eyes in rabbits
eventhough it is not a skin sensitizer or irritant.
It was observed to be a strong respiratory inhibitor in complex I of mitochondria in rat liver
and purified bovine heart.
Fenazaquin inhibits complex I and blocks induction of ornithine decarboxylase by way of
several marketers in human breast cancer cells in vitro. It's far postulated that each the block
of decarboxylase and anti cancer motion arise due to a common reason i.e. the
mitochondrial respiratory chain reduces the production of reactive oxygen species.

 AZOCYCLOTIN (Complex 5 inhibitor):

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 Azocyclotin is a acaricide utilized for cotton, fruits and ornamentals under those exchange
name Peropal. Clinched alongside existing frameworks alternately under basic states
azocyclotin is promptly hydrolysed will yield tricyclohexyltin hydroxide, a related organotin
acaricide. Azocyclotin’s poisonous quality Toward inward breath is especially helterskelter.
Upon purposes of presentation on higher doses indications in summed up misery of health,
passing from claiming weight, sleepiness Also torpidly with looseness of the bowels Also
relaxing issue. It is a solid dermal and destructive eye aggravation. Complex V from
claiming mitochondria is hindered because of the activity of azocyclotin.

 CYAZOFAMID (Complex 3 inhibitor) :

Cyazofamid is far a new systemic foliar and soil fungicide.It has each curative and
preventive functionality. Cyazofamid is said to inhibit complex III but on the
Qisite(antimycin site) which differs from that of different recent fungicidal inhibitors of
complex III which act the Q0 website. It may be guaranteed will make particular in its
inhibitory movement with mitochondria from oomycete growths.

CONCLUSION
Modes of motion of pesticides:
• Disturbance in power creation
• Inhibition of photosynthesis
• Free radical iteration & SH-staff reactivity
• Interference with cell division
• Inhibition of nucleic acid synthesis
• Inhibition of enzymes: Ergosterol synthesis, Amino acid synthesis, Chitin synthesis,
Cholinesterase
• Hormone-like and behaviour-enhancing sellers
Reference:
https://www.khanacademy.org/science/biology/cellular-respiration-and-fermentation/oxidative-
phosphorylation/a/oxidative-phosphorylation-etc
Chemical Pesticides Mode of Action and Toxicology
By Jørgen Stenersen

<Subject Title> 18
Introduction
Liver
The liver is the second biggest organ in the body. It buckles down, performing several perplexing
capacities, including: battling contaminations and disease , expelling poisons (toxic substances, for
example, liquor, from the body controlling cholesterol levels blooding to clump (thicken
discharging bile, a fluid that separates fats and helps processing
Liver needs to perform various types of biochemical, manufactured and excretory capacities, so no
single biochemical test can identify the worldwide elements of liver. All labs ordinarily utilize a
many tests for introductory discovery and administration of liver illnesses and these tests are much
of the time named "Liver capacity tests".
Liver function test and classification
LFT
Liver capacity tests (LFT) are a useful screening instrument, which are a compelling
methodology to recognize hepatic brokenness. Since the liver plays out an assortment of capacities
so no single test is adequate to give finish gauge of capacity of liver. Regularly clinicians are
confronted with reports that don't count with the clinical state of the patient and they confront
trouble in translating the LFT.
A. Tests of the liver’s capacity to transport organic anions and to metabolize drugs- Serum
bilirubin, urine bilirubin, urobilinogen etc.
B. Tests that detect injury to hepatocytes (serum enzyme tests) – Amino transferases, alkaline
phosphatase, glutamyl transpeptidase, 5 nucleotidase, leucine, Aminopeptidase etc.
C. Tests of the Liver’s biosynthetic capacity- Serum proteins, albumin, prealbumin, serum
ceruloplasmin, procollagen III peptide, a 1 antitrypsin, a feto protein, and prothrombin time etc.
LIST OF LIVER FUNCTION TESTS

1) SERUM BILIRUBIN:
Bilirubin is a waste item customarily prepared by the liver. The breakdown of red platelets makes
this waste item. It goes through the liver before being discharged through your stool. A harmed
liver can't appropriately prepare bilirubin. This leads to an unusually abnormal state of bilirubin
into blood. A high outcome on the bilirubin test demonstrates that the liver isn't working
appropriately.
Other conditions in which bilirubin level increases are;
Bilirubin level increases in problems like hemolysis, myoglobinemia, hematoma, ineffective
erythropoiesis .so , peoples with this kind of conditions undergoing serum bilirubin test shows
damage in liver, which is an false positive test regarding liver.

2) ALKALINE PHOSPHATASE TEST(ALP TEST)

alkaline phosphatase (ALP) is an enzyme found in your liver, bile ducts, bone ,muscles. then ALP
test is usually processed with combination with several other tests. if alp test alone processed to
patients with high levels of alp may indicate liver damage.
other conditions in which alkaline phosphatase level increases are:
alkaline phosphatase level increases in like bone diseases, placenta and intestinal problems ,
tumours. so, peoples with this kind of conditions undergoing alkaline phosphatase test(alp test) test
shows damage in liver, which is an false positive test regarding liver.

3) AST(ASPARATE AMINO TRANFERASES)

Aspartate aminotransferase (AST) is an enzyme which seen in several parts of our body. When
these enzymes released into blood stream it shows result on indicate a problem with the liver, it’s
usually measured together with ALT to check for liver problems.

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other conditions in which level Aspartate aminotransferase (ASt) increases are;
Aspartate aminotransferase level increases in condtion like muscle, kidney, bone, cardiac,
brain diseases. so, peoples with this kind of conditions undergoing Aspartate aminotransferase
(AST) test shows damage in liver, which is an false positive test regarding liver. so, doctors
recommend to undergo combination of both AST and ALT.

4) ALT(ALANINE TRANSAMINASE TEST)

Alanine transaminase (ALT) it is used as body metabolizing protein. If ALT level increases it will
lead to release into blood stream which leads and indicates the liver damaged or liver diseases.

other conditions in which ALT(ALANINE TRANSAMINASE TEST) level increases are;

ALANINE TRANSAMINASE Level Increases Conditions Like Thyroid Disorders, Celiac
Diseases, Hemolysis, Muscle Disorders. So, Peoples With This Kind Of Conditions Undergoing
ALANINE TRANSAMINASE (ALT) Test Shows Damage In Liver, Which Is An False Positive
Test Regarding Liver

5)Prothrombin time
Prothrombin is a protein delivered by your liver. It is one of many calculates your blood that help
it to clump fittingly. On the off chance that you take a blood-diminishing drug, for example,
warfarin (Coumadin), normal prothrombin time test results will be Normal control usually is in the
range of 9-11seconds. A prolongation of more than 2 seconds is considered abnormal so , low
outcome on this time demonstrates that your liver isn't working appropriately.
other conditions in which Prothrombin time increases are seen in conditions vitamin
deficiency secondary IN PEM,MAS,DIC, various deficiencies of coagulation factors, DIC,and
ingestion of certain drugs.so, peoples with this kind of conditions undergoing Prothrombin time
test shows damage in liver, which is an false positive test regarding liver.

Conclusion : A liver capacity test is of little esteem in screening for liver sickness the same
number of genuine liver maladies might be connected with ordinary levels and irregular levels may
be found in asymptomatic sound people. The utilization of battery of liver capacity tests, however
constitutes a profoundly delicate method. The quantity of false negatives must be diminished by
this procedure. The utilization of battery of liver tests is additionally connected with high specificity
particularly when more than one test is irregular. Since the liver performs a variety of functions so
no single test is sufficient to provide complete estimate of function of liver.
References:
http://www.healthline.com/health/liver-function-tests#Overview1
http://medind.nic.in/icb/t07/i7/icbt07i7p663.pdf
http://www.aafp.org/afp/2011/1101/p1003.html

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