Lab practical report on iron chloride detection of epinephrine hormone

1. KENYATTA UNIVERSITY
SCHOOL OF MEDICINE.
DEPARTMENT OF MEDICAL BIOCHEMESTRY
NAME: LANDO ELVIS OTIENO
REG NO: P29S/16344/2015.
COURSE: MBCHB
LECTURER: Dr STEPHEN NYANCHOMA
HANDING DATE: 18 /4/2017
SUBJECT: PRACTICAL REPPORT ON DETECTION OF EPINEPHRINE
HORMONE WITH IRON CHLORIDE.
ELVIS L
©

2. INTRODUCTION
Epinephrine(Ep),oftencalledadrenaline,isone of the mostimportantneurotransmittersinmammalian
central nervous systems, existing in the nervous tissue and body fluid, and controlling the nervous
system in the performance of a series of biological reactions and nervous system chemical processes.
Many biological reactions, and the nervous system chemical processes can be considered as a type of
organicelectrochemical process. Therefore,astudyonthe electrochemical behaviorof Ep isa great help
inunderstandingthe effectsof the nervoussystemandartificial substitutes, and is of great significance
to pharmacological research and the life sciences. Determination of concentrations of Ep is important
for the studyof neurotransmission,fordiagnosisof neurological disorders, such as Parkinson's, and for
developing medicines to treat the diseases
Epinephrine may be quantified in blood, plasma or serum as a diagnostic aid, to monitor therapeutic
administration, or to identify the causative agent in a potential poisoning victim. Endogenous plasma
epinephrine concentrationsinrestingadultsare normallylessthan10 ng/L, but may increase by 10-fold
duringexercise andby50-foldor more during times of stress. Pheochromocytoma patients often have
plasma adrenaline levels of 1000–10,000 ng/L. Parenteral administration of epinephrine to acute-care
cardiac patients can produce plasma concentrations of 10,000 to 100,000 ng/L
PRINCIPLE
Epinephrine( adrenaline, N-methylaminomethanolpyrocatechol) may be determined by reactions
characteristic for pyrocatechols. It forms phenolate-like compound which has green color in the
presence of ironions.Epinephrine iseasilyoxidizedthat also is characteristic for pyrocatechols, if alkali
isaddedto epinephrine solutionorto its complex with iron, red colored adenochrome will be formed.
REAGENTS AND MATERIALS
-0.1% epinephrine solution(sample one)
-1% FeCL3 solution
-10% NaOH
-control(sample 2)

3. -test tubes
-dropper
PROCEDURE
Take 10 drops of 0.1% epinephrine(adrenaline) solution into a test tube and add 1 drop of 1% FeCL3
solution. Blue green color appears.
After that add 1 drop of 10% NaOH to the received solution. The solution of the liquid becomes dark
red.
OBSERVATION AND RESULT
In sample one the color of the solution turned to Blue green color on addition of 1 drop of 1% FeCL3
solution. Which inturn changed color to dark red on further addition of 1 drop of 10% NaOH.
In sample two(2),there wasnocolorchange on additionof 1 drop of 1% FeCL3 , however on addition of
1 drop of 10% NaOH, the solution became rusty in color( egg green)
Therefore sample one solution waspositiveforpositive forepinephrinetest,andsample 2wasnegative
for epinephrine( acts as a control).
DISCUSION AND CONCLUSION
The adrenal medulla is a minor contributor to total circulating catecholamines (L-DOPA is at a higher
concentration in the plasma), though it contributes over 90% of circulating epinephrine. Little
epinephrine is found in other tissues, mostly in scattered chromaffin cells. Following adrenalectomy,
epinephrine disappears below the detection limit in the blood stream.
The adrenals contribute about 7% of circulating norepinephrine, most of which is a spill over
from neurotransmission with little activity as a hormone.Pharmacological doses of epinephrine
stimulate α1, α2, β1, β2, and β3 adrenoceptors of the sympathetic nervous system. Sympathetic nerve
receptors are classified as adrenergic, based on their responsiveness to adrenaline.
During exercise the epinephrine blood concentration rises partially from increased secretion from the
adrenal medullaandpartlyfromdecreasedmetabolismbecauseof reducedhepaticbloodflow. Infusion
of epinephrine to reproduce exercise circulating concentrations of epinephrine in subjects at rest has
little haemodynamiceffect,other than a small β2-mediated fall in diastolic blood pressure. Infusion of
epinephrine wellwithinthe physiological range suppresseshumanairwayhyper-reactivitysufficientlyto
antagonize the constrictor effects of inhaled histamine
effectson memoryand emotion: Ithas beenfoundthatadrenergichormones,suchasepinephrine, can
produce retrograde enhancementof long-term memory in humans. The release of epinephrine due to
emotionallystressful events,whichisendogenousepinephrine,canmodulate memory consolidation of
the events, ensuring memory strength that is proportional to memory importance. Post-learning

4. epinephrine activity also interacts with the degree of arousal associated with the initial coding.
Epinephrinedoesnotreadilycrossthe blood–brainbarrier,soitseffects onmemoryconsolidationare at
least partly initiated by β adrenoceptors in the periphery.
Mode of action: As a hormone, epinephrine acts on nearly all body tissues. Its actions vary by tissue
type and tissue expression of adrenergic receptors. For example, high levels of epinephrine
causessmoothmuscle relaxationinthe airwaysbut causes contraction of the smooth muscle that lines
most arterioles.
Epinephrineactsbybindingtoa varietyof adrenergicreceptors.Epinephrine isanonselectiveagonistof
all adrenergic receptors, including the major subtypes α1, α2, β1, β2, and β3. Epinephrine's binding to
these receptors triggers a number of metabolic changes. Binding to α-adrenergic receptors
inhibits insulin secretion by the pancreas, stimulates glycogenolysis in the liver and muscle, and
stimulates glycolysis andinhibitsinsulin-mediated glycogenesisinmuscle.βadrenergic receptor binding
triggersglucagonsecretioninthe pancreas,increased adrenocorticotropichormone (ACTH) secretionby
the pituitary gland, and increased lipolysis by adipose tissue. Together, these effects lead to
increased blood glucose and fatty acids, providing substrates for energy production within cells
throughout the body.
Its actions are to increase peripheral resistance via α1 receptor-dependent vasoconstriction and to
increase cardiac output via its binding to β1 receptors. The goal of reducing peripheral circulation is to
increase coronary and cerebral perfusion pressures and therefore increase oxygen exchange at the
cellular level. While epinephrine does increase aortic, cerebral, and carotid circulation pressure, it
lowers carotid blood flow and end-tidal CO2 or ETCO2 levels. It appears that epinephrine may be
improvingmacrocirculationatthe expense of the capillary beds where actual perfusion is taking place
Biosynthesis: In chemical terms, epinephrine is one of a group of monoamines called
the catecholamines.Itisproducedinsome neurons of the central nervoussystem,andinthe chromaffin
cells of the adrenal medulla from the amino acids phenylalanine and tyrosine.[67]
Epinephrine is synthesized in the medulla of the adrenal gland in an enzymatic pathway that converts
the amino acid tyrosine into a series of intermediates and, ultimately, epinephrine. Tyrosine is first
oxidized to L-DOPA, which is subsequently decarboxylated to give dopamine. Oxidation
gives norepinephrine. The final step in epinephrine biosynthesis is the methylation of the primary
amine of noradrenaline. This reaction is catalyzed by the enzyme phenylethanolamine N-
methyltransferase (PNMT) which utilizes S-adenosyl methionine (SAMe) as the methyl donor. While
PNMT is found primarily in the cytosol of the endocrine cells of the adrenal medulla (also known
as chromaffin cells), it has been detected at low levels in both the heart and brain.
Regulation:The majorphysiologic triggers of adrenaline release center upon stresses, such as physical
threat,excitement,noise,brightlights,andhighambienttemperature.Allof these stimuli are processed
in the central nervous system.

5. Adrenocorticotropic hormone (ACTH) and the sympathetic nervous system stimulate the synthesis of
adrenaline precursors by enhancing the activity of tyrosine hydroxylase and dopamine β-hydroxylase
Unlike many other hormones adrenaline (as with other catecholamines) does not exert negative
feedback to down-regulate its own synthesis. Abnormally elevated levels of adrenaline can occur in a
varietyof conditions,suchassurreptitiousepinephrine administration, pheochromocytoma, and other
tumors of the sympathetic ganglia.
Its action is terminated with reuptake into nerve terminal endings, some minute dilution, and
metabolism by monoamine oxidase and catechol-O-methyl transferase.
Clinical application:As a medication epinephrine is used to treat a number of conditions,
including anaphylaxis, cardiac arrest, and superficial bleeding. Inhaled epinephrine may be used to
improve the symptoms of croup. It may also be used for asthma when other treatments are not
effective.Itisgiven intravenously,byinjectionintoamuscle,byinhalation,orbyinjectionjustunderthe
skin. Common side effects include shakiness, anxiety, and sweating. A fast heart rate and high blood
pressure mayoccur. Occasionallyitmayresultin an abnormal heart rhythm. While the safety of its use
duringpregnancyandbreastfeeding is unclear, the benefits to the mother must be taken into account
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