o Heme is incorporated in HgB which

carries oxygen
TRACE ELEMENTS AND PORPHYRINS • Hormonal activities
o Different hormones in the body which
Week 1: Lecture Discussion
may be produced in the presence of
trace elements
TRACE ELEMENTS • Enzyme activator or cofactors
• Found in the body in very little concentration o Coenzyme: organic compounds
o Known to constitute less than 0.01% of o Cofactor: inorganic compounds
the total body weight because our total o Without trace elements, coenzymes/
body water (TBW) is composed of 96% cofactors cannot work. TE activates
water 3.98% electrolytes and the cofactor/ coenzyme
remaining of the 100% are made up of • Protection from free radicals
trace elements o SOD (Superoxide dismutase): enzyme
• Essential or Non-essential composed of zinc and copper; has
o Essential: if a deficiency of an element antioxidant effect; known to be
impairs biochemical or functional scavenger of free radicals
processes and replacement of it ▪ Free radicals present (2O2) →
corrects impairment reacts with Superoxide
▪ Cannot be synthesized by the dismutase→ (2O2) is converted
body in a large amount (Can to O2 + H2O2
be synthesized but minute/ ▪ H2O2 → reacted with catalase
limited amount only) or peroxidase → Producing
• Therefore, should be H2O + O2
present in our diet o Selenium: known component of
• CAUSES of decreased glutathione, and regenerates Vit. D and
trace elements in the C
body: Decreased • Tissue repair (Zn)
intake, impaired o Zinc sulfate: Active agent of wound
absorption. Increased healers
excretion, genetic • Immune system
abnormalities • Growth and Reproductive process
▪ Metalloenzymes/
Metalloproteins- enzymes/
proteins that work together
IRON
with the trace elements • Most abundant trace element present in the
▪ Organ malfunction if trace body
elements are deficient, or may • Present in the body in two states:
cause impairment of o Ferric (Fe3+)
biochemical processes o Ferrous (Fe2+)
o Non-essential: may be toxic in the body • Constituent of hemoglobin and myoglobin
if increased in amount o Needed for heme synthesis
• Toxicity- excess concentration of trace element o Hemoglobin
which may lead to intoxication ▪ Hgb carrier of oxygen (iron is
found in heme part of
10 Essential Trace Elements: hemoglobin, basically the one
that carries the oxygen in the
1. Copper- 3rd most abundant
Hgb)
2. Chromium
o Myoglobin- protein for muscles
3. Cobalt
▪ Serves as an agent in redox
4. Iron- 1st most abundant
reaction and electron transport
5. Manganese
• Cofactor: cytochrome oxidase, xanthine
6. Molybdinum
oxidase, peroxidase, catalase
7. Zinc- 2nd most abundant
• Absorbed in the duodenum and jejunum
8. Selenium
o Iron has 2 states, ferric and ferrous
9. Iodine
o Ferric is the form of iron found in our
10. Fluorine
food/ diet. The food with ferric will go to
➔ Halogen
stomach (has acidic pH which will
convert ferric to ferrous)
Biologic Functions
▪ May be converted from ferric
• Electron transport to ferrous by Vitamins C and/or
• Respiration (Iron) acidic environment of stomach
o Needed for heme production
 o Ferrous (absorbable form) form can • Apoferritin is absent - iron will be stored as
now go to small intestine → Duodenum hemosiderin
→ Jejunum (where absorption of iron o Cases of inflammation/infection –
happens) causes a decrease in apoferritin
▪ Transferrin and albumin will production in the liver, bone marrow,
transport iron to different orans/ and the spleen; iron will be stored as
parts of the body (Mainly, bone hemosiderin.
marrow, liver, spleen- where • Source of apoferritin - Hepatocytes of the liver,
iron is stored as ferritin or bone marrow, spleen, and macrophage.
hemosiderin) • Tissue that contains Hemosiderin
Absorption • Hemosiderin
• To be functional and absorbed by the small • There is a decrease in the production of
intestine (duodenum & jejunum) it should be in apoferritin
ferrous form: o A small oxide granule that can be
o Ferric (iron in diet) → acidic pH of stained by prussian blue – hemosiderin
stomach (contains HCl, organic acid, will look like blue to black granules.
vit. C and enzyme ferric reductase) → o Iron storage in the form of hemosiderin
converted to ferrous → absorbed by o Another form of extracellular iron can
duodenum and jejunum → complex to also be stained by pearl stain.
transferrin & albumin → goes to bone • Over stimulation of macrophages
marrow, liver, spleen (where iron is o This is an increase in apoferritin
stored as hemosiderin or ferritin) production.
o Presence of increased apoferritin in the
• Transport Proteins: Transferrin/Albumin circulation.
• Stored as Ferritin or Hemosiderin o More iron will bind to the apoferritin
o Ferritin: composed of 4500 atoms of iron ▪ Increase in apoferritin will
▪ Ferritin = iron + apoferritin cause an increase in iron
(source: bone marrow, liver, ▪ Increased in formed ferritin
spleen, macrophages) ▪ Result to a decrease in
▪ If there is no production of circulating iron in the blood or
apoferritin, iron will be stored as cause deficiency for iron
hemosiderin which happens in • Remember that ferritin
cases of inflammation/ is for storage not for
infection stimulation
▪ Over stimulation of
macrophages will lead to an Hemoglobin molecule
increased production of both • Heme - contains the
apoferritin and ferritin and a iron that carries the oxygen
decreased level of iron in the molecule
blood • Globin
o Hemosiderin – extracellular form of iron
which can be stained using pearl stain
▪ Not the primary storage form of Iron Overload
iron
→ Excess iron in plasma is toxic which results to pro-
▪ Temporary storage of iron
oxidation of cells, and as a result is the
when apoferritin is absent
destruction of DNA
▪ Can be stained by crucial blue
→ Cannot be excreted in urine or stool
or pearl stain
→ Can only be excreted through menstruation (for
• Accumulation of free Iron in the plasma is toxic
female)
o When absorbed it should be
→ AKA Hemosiderrhosis
transported to organs of the body that
Causes:
it can be stored
• Increased RBC destruction
o Hemolytic anemia
• A storage form
o Lysed RBC releases Hgb which contains
of iron → Ferritin
iron
(Ferritin is made up of
• Increased mobilization of Iron
apoferritin)
o Increase mobilization of free iron in case
o Oxidized to be
of an increase of ceruloplasmin in
in the ferric state so
blood
that apoferritin can
▪ A protein responsible of the
bind with it
transport of copper
▪ Apoferritin + Iron = Ferritin
▪ Known to facilitate iron
mobilization from ferritin
 ▪ Increased ceruloplasmin is • Early morning sample (12 hrs. fasting)
present in hepatic cirrhosis o Diurnal variation
• Decreased blood cell formation o Increases in the morning
o Failure of iron to form heme or o Decreases in the afternoon
protoporphyrin • Measures Free iron (present in the body)
o Lead poisoning, thalassemia, Vitamin • Normal Values:
B16 deficiency o Males: 65-177 ug/dL (11.6-31.7 umol/L)
• Defective Iron storage o Females: 50-170 ug/dL (9.0-30.4 umol/L)
o Pernicious anemia
Fe Fe II Ferren
o Iron cannot be absorbed by the small Fe III
separated complexed e
reduced to
intestine from binding
Fe II
with Terosit
• Increase rate of absorption proteins chromogens e
,
o Results to Hemosiderosis
o Or due to repeated blood transfusion METHOD:
• Collectively referred to as hereditary • Iron should be separated first from its binding
hemochromatosis proteins through the use of a strong acid (do not
o Accumulation of iron in important use trichloroacetic acid or TCA)
organs of the body (like endocrine • Ferric will be reduced to ferrous form through the
glands, liver, heart, spleen) use of reducing agent such as ascorbic acid or
Differentiating Hemochromatosis VS Iron Overload any acidic substance

→ Iron overload/Hemosiderosis
1. Fe separated from binding proteins (transferrin and
• First thing that happens
albumin) using a strong acid except trichloroacetic acid
• Will continue to happen in case of repetitive
2. Fe III (ferric state) reduced to Fe II (ferrous state) using
blood transfusion and will eventually lead to
a reducing agent (i.e. ascorbic acid), but other acidic
hereditary hemochromatosis
substances can also be used as a reducing agent.
o There will be accumulation of iron in the
3. Fe II (ferrous form) Complexed with chromogens
liver, heart, spleen, and important
endocrine glands
→ Product is measured spectrophotometrically
o May cause the organs to malfunction
Chromogens:
• Hereditary hemochromatosis → Malfunctioning
• Ferrene
• of organs due to repetitive transfusion.
• Terosite
• Hemosiderosis/Iron overload → *Just* iron
• 2,4,6-Tripyridyl S-Triazine
overload
• Sulfobathophenanthroline (SBP)

→ Stored iron goes to the bone marrow for

→ End color is being measured through
hematopoiesis
spectrophotometry
→ Excess iron in the plasma
→ Toxic because it can result to peroxidation of cells
Total Iron Binding Capacity (TIBC)
resulting to the destruction of the DNA
→ Iron should be excreted • Indirect measure of serum transferrin or free
• It can be during menstruation transferrin
• Amount of iron that could be found by
saturating the binding proteins present in the
Iron Deficiency
sample is being measured
→ Decrease iron in plasma • Differentiates IDA from anemia of chronic
disease
• Generalized Iron deficiency o IDA:  TIBC (Increased TIBC)
o IDA o ACD:  TIBC (Decreased TIBC)
• Chronic loss of iron • Due to overstimulation of macrophage, which
o hemorrhagia results to decrease iron level in the blood
• Impaired release of Iron from RES o Increased macrophage = decreased
o Impairment with mobilization of iron due iron level
to decrease ceruloplasmin • Normal Values: 240-450 ug/dL (43.0-80.6 umol/L)

• Signs & symptoms: Percent Saturation

o Fatigue & tiredness, shortness of breath, • Measures % transferrin bound with iron
depression, hair loss, leg syndrome, • Normal Values: 20-55%
frequent headache, increase sensitivity • FORMULA- % Saturation = (Serum Fe/TIBC) x 100
to cold, brittle nails • Differentiates IDA from anemia of chronic
disease
Laboratory Evaluation of Iron o IDA:  Percent saturation
Serum Iron o ACD:  Percent saturation

• Non-hemolyzed serum
 ZINC
Serum Transferrin
• Component of >300 enzymes
• Direct measurement of transferrin
o Helps enzyme do its function
• Measured using nephelometry assay
▪ Needs iron
• An index of iron nutritional status
o Serves as cofactor of enzymes
• Measures free transferrin
• Absorption is through metallothionine
• Differentiates IDA from anemia of chronic
o Shares same absorption process with
disease (sobrang daming iron sa body kaya
copper
mataas din value ng transferrin)
• Albumin and alpha-2-macroglobulin
o IDA:  Serum transferrin
o Transports zinc
o ACD:  Serum transferrin
• Nutrient
• Normal Values: 200-400 mg/dL o Promotes healthy immune system
• Serum transferrin: TIBC (ug/dL) x 0.70 = mg/dL o Promotes wound healing
o Promotes ovulation, sperm
Serum Ferritin
development
• ELISA, Chemiluminescence o DNA synthesis, growth development
• An immunoassay o Metabolism of protein, glucose and
• An index of iron stores (storage form of iron) cholesterol
• Differentiates IDA from anemia of chronic • Second most abundant
disease • Increase Zinc MEANS decrease circulating
o IDA:  serum ferritin copper
o ACD: serum ferritin Zinc excess
→ Serum transferrin () and ferritin () results of ACD are
the same with iron overload and hereditary • Intoxication is rare
hemochromatosis o Zinc is mostly considered as nutrient
Serum Iron Iron storage • Zinc Fume Fever
ferritin concentration o There is an increase in zinc present in the
in blood
body because of zinc oxide fumes and
ACD Inc Dec Inc
dust that could be inhaled resulting to
IDA Dec Inc Dec
TIBC Transferrin Ferritin % Con. Of pneumonia in the lungs and that is one
Saturation iron known effect of an increasing presence
(plasma)
in the plasma or the serum of an
ACD Dec Dec N/ Inc Dec Dec
individual or a patient
IDA Inc Inc Dec Dec Dec

SUMMARY: Zinc Deficiency

● IDA • Growth retardation
→ Increased TIBC • Delayed sexual maturation
→ Increased Transferrin o Testicular atrophy
→ Decreased Ferritin • Prolonged wound healing
→ Decreased percent saturation o Acrodermatitis enterophatica
→ Decreased in the concentration of iron present • Impaired wound healing
in body or in the plasma
● ACD Acrodermatitis enterophatica
→ Decreased TIBC • There’s a problem in zinc reabsorption
→ Decreased Transferrin • Group of disorder characterized by dermatitis,
→ Increased Ferritin / Iron storage hyperpigmented lesions, alopecia, growth
→ Decreased percent saturation retardation, infection, diaper rash, and facial
→ Decreased in the concentration of iron present rashes
in body or in the plasma
COPPER
• Metalloenzymes and proteins
o Metalloenzymes: ceruloplasmin,
cytochrome C oxidase, SOD, tyrosinase,
lysyl oxidase, dopamine hydroxylase,
CF V, metallothionine
o Critical for reduction of iron in heme
synthesis
• Cofactor of many metalloenzymes and your
proteins
• A component of your SOD together with your
zinc that serves as an antioxidant
o It assumes antioxidant activity with your
Superoxide Dismutase (SOD) and that
 SOD could be present in your Explanation: How is copper being converted and the
erythrocuprein inside your RBCs goal to the different organs in the body especially the
• Copper is also known for having excellent liver?
electrical and heat conductivity and Same as • Copper is now complex with your albumin
your iron, copper has four oxidation states (Cu, because it is now reabsorbed by the stomach or
Cu+1, Cu+2, Cu+3). Among these four-oxidation the small intestine and it will now enter the liver
state, Cu+2 is the most stable of all • In the liver, we have three phases of your
• Constituent of erythrocuprein copper,
• Intestinal absorption o it either binds with your
• Copper is absorbed in small intestine in its apoceruloplasmin
oxidized form o it can go to the biliary tract as a biliary
o The copper that we ingest from the diet copper and go to the bile canaliculus
should be oxidized and when it is o Can be stored as metallothionein which
oxidized, it can now be absorbed in the is a non-toxic form of copper
stomach and in the small intestine • However, when it binds with your
o When it is absorbed, it can now go apoceruloplasmin, that’s the time it becomes a
through the blood and from the blood, ceruloplasmin, and then this ceruloplasmin is
it will be transported to other parts of bounded to your six copper atoms and when it
the body by means of transport protein is now then again in its ceruloplasmin form, it can
(Transcuprein, albumin, and go out of the liver and go back to the blood
ceruloplasmin). circulation where it can now be transported or
o When it is bounded to transport protein, distributed all throughout the body
it can now be transported to different
parts of the body especially the liver Notes
o From the liver, it can now enter the • Ceruloplasmin is bounded by six copper atoms
circulation but this time as • 90% of the copper on the body is in its
ceruloplasmin ceruloplasmin form
• Ceruloplasmin
o Protein that can transport your copper BIOLOGICAL FUNCTION: Copper
however ceruloplasmin is somehow also
• Antioxidant defense
a storage form of your copper and
o Known component of your SOD
once it is incorporated into your
• Collagen formation
ceruloplasmin, it can now be distributed
o Lysyloxidase
all throughout the body
o Serves as a cofactor of your lysyloxidase
• Transcuprein, albumin, and ceruloplasmin
o Lysyloxidase is an enzyme important for
o Known transport protein of your copper
cross linking the protein into a larger
• Metallothionine
fiber
o Unknown enzyme that crosslinks
• Melanin production
carotene in hair that also uses copper
o Tyrosinase
as cofactor
o Serves as a cofactor of tyrosinase
o Without cofactor, enzyme cannot
important for melanin production
perform fully its function
• Production of neurotransmitters
• Copper is also critical for reduction of iron in
o Norepinephrine
heme synthesis
o B-dopamine monooxygenase
• In our stomach there’s a presence of so-called
• Serves as cofactor of your metallothionine
histidine gluconate and citrate that can
• Excretion of copper is through first bowel
enhance absorption of your copper
because it can go to the biliary tract and can
be excreted in the bowel
o 98% of excretion happens in that
manner
o can be excreted also through urine,
sweat, menstruation, and same as for
your iron
• Copper has the most important biologic
functions in the body because it is unknown
cofactor of many metalloenzymes, important in
the different processes that happens in our body

COPPER EXCESS
• Excess copper actually binds with the thionine
producing a metallothionine wherein
 metallothionine is the storage form of your • Severe manifestation: mental retardation
copper and it is a non-toxic • Connective tissue defects
• 98% of copper is excreted through the bowel Signs and Symptoms:
because it can enter the biliary tract outside the • Hypotonia
bile canaliculus and it can mix up with bile and o Muscles are affected
excreted in there. This is how we prevent copper o Disturbance of the muscle tone
excess, however, there are clinical conditions • Mental retardation
that can have this copper excess present in the • Tissue defects
body (Example: Wilson’s disease) • Bones are affected
• Hypothermia
WILSON’S DISEASE • Seizures
o Known excess copper present in the body/
plasma/ circulation Increase zinc absorption
o Known as hepatolenticular degeneration • Zinc and copper somehow share the same
wherein in here, we could see the clinical absorption matter
signs and symptoms includes this Kayser- • If there is a decreased copper because of
Fleischer ring present in the eyes decreased absorption or depending on the
reason that results to an increasing absorption
Manifestations: • Increased Zinc absorption will result in copper
• Increase urinary excretion of copper deficiency
• Decrease ceruloplasmin o High dose of zinc supplement =
• Kayser-Fleischer ring decrease copper absorption = copper
• Increase metallothionine deficiency
o Storage form of copper • Treatment for Wilson’s disease
o Will lead to high levels of copper in the
body
CHROMIUM
Medications: • Comes from the Greek word “chroma” meaning
• D-penicillamine, Dimercaprol, Ammonium color
tetramolybdate • Potentiates insulin action
o Drugs given to those patients suffering o Chromium is used as a supplement for
from Wilson’s disease diabetic patients
o Inhibits copper absorption in the o Promotes carbohydrate metabolism as
stomach in the small intestine well as fats in cholesterol metabolism
o D-penicillamine and Dimercaprol: are • Glucose tolerance factor
known to be promoters of urinary • Chromium levels in hair is higher than serum
excretion of copper • Brewer’s yeast
o Ammonium tetramolybdate: inhibits o Saccharomyces cerevisiae is a good
copper absorption in the stomach and source of chromium
the small intestine Note:
• Zinc • Chromium excess or deficient is also present in
o High dose of zinc supplement = the body but it’s not somehow of medical
decrease copper absorption = copper interest, it’s not abundant in the body
deficiency
FLUORINE
Signs and Symptoms: • Incorporated into bone crystals
• Prevents occurrence of osteoporosis and
somehow it has the same action as your calcium
• Corrects calcium deficiency
o Fluorine is related to the action of
calcium
o Whenever there’s calcium deficiency,
fluorine can be of great used to correct
that deficiency
• Enhanced bone formation
COPPER DEFICIENCY o Prevents occurrence of osteoporosis
• Replace calcium in increase bone density
MENKE’S KINKY HAIR SYNDROME • Excess fluorine: result to discoloration of enamel
in teeth
• Defects in copper transport
o When you use a toothpaste that
• It is a known connective tissue and sex-linked
contains too much fluorine
defect
concentration this will result to a pitted
Manifestations:
teeth or crack teeth or discoloration
• Results to “Kinky/ steely hair”
 COBALT ▪ epilepsy, hip abnormalities,
joint diseases, congenital
• Component of Vitamin B12
malformation, heart and bone
o Whatever is the function of Vit B2 is
problems and stunted growth
important for the cobalt
in case of children
o Without cobalt, there’s no Vit B12 and if
there’s no Vit B12, the function of the Vit MOLYBDENUM
B12 will not be executed
• Basis of treatment of Wilson’s Disease
• Absorbed by the same mechanism as Iron
o Disease with excess copper in the blood
o Heme incorporated with iron = Cobalt
• Inhibits Copper and Iron absorption
incorporated with Vit B12
• Xanthine oxidase, xanthine dehydrogenase,
o Heme and Vit B12 has the same
sulfite oxidase, aldehyde oxidase
Structure
o Known enzymes that uses molybdenum
MANGANESE as their component/ cofactor
o Molybdopterin
• Unknown metalloenzymes constituent and a
▪ Known cofactor
very known enzyme activator
o The active site of these following
• It is common nowadays because it is commonly
enzymes will bind to molybdenum
used for steel production
producing molybdopterin
• High dosage is NOT toxic except when inhaled
Notes:
o Acute Manganese Aerosol Intoxication:
• Many of the trace element assumes as a
Locura manganica (Manganese
component or as a coenzyme of the different
madness)
enzymes in the body
▪ Common in miners when they
• Molybdenum deficiency is rare
inhaled too much manganese.
• Molybdenum cofactor deficiency is somehow a
During mining there’s an
known inherited condition
increased manganese content
• Molybdenum toxicity is also rare
that could be present in the
• Few known cases of exposure to human are
environment
reported
• MRI patients: has Manganese deposition in the
• Whenever there’s exposure, there’s a recorded
brain
increase in the uric acid and increase incidence
o The dye used for MRI is composed of
of gout to the individual
Manganese
• Pyruvate carboxylase, mitochondrial SOD
(Superoxide dismutase), arginase and SELENIUM
glucokinase
• Once considered as carcinogen
o These are enzymes that is known to use
• Naturally present in the soil
Manganese as their coenzyme
• Regenerates Vitamin E And Vitamin C which are
together with iron and copper
known as the most potent vitamins to fight
o Copper is mostly used or the first choice
cancer cell formation
for metalloenzymes however, it can be
• A known component of Glutathione
replaced by Manganese or vice versa
peroxidase:
as an activator of this enzyme masking
o Selenocysteine
the effect of the deficiency just in case
o Part of cellular antioxidant defense
deficient in manganese or copper
mechanism against free radicals
• Chronic manganese toxicity: Observed in
• Can be involved in the metabolism of thyroid
Parkinson’s disease
hormones namely T3 and T4
o Akinesia, Rigidity, Tremors, and mask-life
• Good sources of selenium are:
faces
o Root crops and vegetables
• Manganese Toxicity
• Selenium Excess
o Liver disease
o Acute oral exposure- manifested in GIT
▪ Increased manganese present
and cardiovascular symptoms (nausea,
in the body or in the plasma
vomiting, diarrhea, tachycardia)
resulting to a decreased
o Chronic oral exposure- dermal effects
manganese excretion or
and neurological problems (skin and
elimination
neurologic problems)
▪ Promotes accumulation of
• Selenium deficiency
manganese in various region of
o Keshan Disease
the brain that could be result in
▪ Endemic myocardiopathy
mental retardation
▪ Presented they have a juvenile
• Manganese Deficiency:
cardiomyopathy
o Low levels of Manganese have been
▪ Destruction of cardiac cells
associated with
 o Kashin-Beck Disease fluid, it denotes an abnormality
▪ Endemic osteoarthritis in heme synthesis
▪ Chondronecrosis is necrosis in ▪ Porphyrin is important for the
the cartilage formation of the heme ring
Notes:
• Deficiency means decreased level of trace • PORPHYRIAS
elements in the blood. Either bounded in their o Clinical conditions related to porphyrins
transport protein o Aberration/Disturbances in heme
• Excess or overload- increased level of trace synthesis
elements in the blood o Accumulation of Porphyrins
• Most of the trace elements are known to be a ▪ Depending on the specimen
component of a coenzyme and form
• Abundancy/Three commonly measured in the o Classifications:
laboratory: ▪ Acquired
o Iron → most requested by doctors ▪ Inherited
o Zinc • Porphyrinogens
o Copper o Reduced form of porphyrins
o Most common porphyrins present in the
body
PORPHYRINS o Almost all porphyrins present in the
body is in the form of porphyrinogens
• Came from the heme ring of the hemoglobin o Considered as the functional form of
portion the compound that must be used
o Together with hemoglobin, myoglobin during heme synthesis
contains porphyrin groups
▪ Both hemoglobin and Porphyrins VS Porphyrinogens
myoglobin are chemically Characteristics Porphyrinogens Porphyrins
similar in molecules that Stability Unstable Stable (uro,
contain porphyrin groups copro, proto)
o When iron is escalated from the Fluorescence Do not fluoresce DOES fluoresce
porphyrins, the heme ring is formed (colorless) (produces
▪ When heme ring binds with characteristic
globin portion (protein), colors)
hemoglobin is formed in the
body • When porphyrinogens are oxidized to
o Hemoglobin, when degraded, porphyrins, upon exposure to air, it produces the
produces heme and globin portions characteristic color
(protein portion of hemoglobin)
PORPHYRINS (HEME SYNTHESIS) PATHWAY
• Porphyrins: Metabolic intermediates in the
Hemoglobin synthesis
o Important in hemoglobin synthesis
o Myoglobin and respiratory pigments
(cytochromes) also contain porphyrins
o Result: hemoproteins (hemoglobin)
• Red violet to red-brown in color
o Produced in vitro
• THREE FORMS OF PORPHYRINS: URO, COPRO,
and PROTO
o 3 forms to measure or are present in
biological fluid
o URO → uroporphyrin
▪ Found in urine
▪ Most soluble in water
o COPRO → coproporphyrin
▪ Found in urine, stool, and
Notes
sometimes in blood
• Main site of heme synthesis is the liver and bone
o PROTO → protoporphyrin
marrow
▪ Found in stool
• Reduction in enzyme present in the pathway
o NOTE: The three forms are considered to
results in either excess production of either one
be clinically-significant porphyrins in
or more precursor substances
humans
• Deficiency of enzymes are the main reason or
▪ Reason: When the porphyrins
cause as to why there are porphyrias
are increased in biological
 Explanation of Pathway • Uroporphyrinogen-III is acted upon by
Uroporphyrinogen decarboxylase (UROD)
Glycine + Succinyl-CoA
o Product: Coproporphyrinogen-III

Coproporphyrinogen-III

• Raw materials for heme synthesis

• Acted upon by ALA synthase
o Product: δ (Alpha)-Aminolevulinic acid • Coproporphyrinogen-III is acted upon by
(δ-ALA) Coproporphyrinogen oxidase
o Product: Protoporphyrinogen-III
δ-Aminolevulinic acid
Protoporphyrinogen-III

• δ-ALA is acted upon by ALA dehydratase or ALA

synthase (first enzyme)
• Protoporphyrinogen-III is acted upon by
o Product: Porphobilinogen (PBG)
Protoporphyrinogen oxidase
▪ PBG is the first precursor
o Product: Protoporphyrin-III
substance involved in the
heme synthesis pathway
Protoporphyrin-III
Porphobilinogen

• PBG is acted upon by Uroporphyrinogen-I • Protoporphyrin III is acted upon by

synthase or hydroxymethylene synthase Ferrochelatase (heme synthase)
(second enzyme) o Product: Heme
o Product: Hydroxymethylbilane or
Uroporphyrinogen-I (linear tetrapyrrol) Additional Notes
• Heme synthesis happens in the mitochondria
Hydroxymethylbilane • Other pathways happen in the cytoplasm or
cytosome
• If there are missing or deficient enzyme in the
pathway, there will be delays in the heme
synthesis
o There is excess production or
accumulation of one or more precursor
substances → porphyria
• Hydroxymethylbilane or Uroporphyrinogen-I is • Overproduction of heme
acted upon by either Uroporphyrinogen-III o Overactive enzyme
cosynthase o Have an effect on the organ and the
• or there is a spontaneous production of main site of heme synthesis.
Uroporphyrinogen-I o May result to porphyria II
o Product: Uroporphyrinogen-III o Overproduction may result to
▪ Accumulation
Uroporphyrinogen-III ▪ Excretion of toxic precursor
compounds including
porphyrins.
• REMEMBER: Overproduction of the precursor
and deficiency in enzymes involved may result
in porphyrias.

PORPHYRIAS
• PP - Plumboporphyria
• AIP - Acute intermittent Porphyria CUTANEOUS SYMPTOMS
• CEP - Congenital Erythropoietic Porphyria
• Photosensitivity
• PCT - Porphyria Cutanea Tarda
o Usual presenting symptom of porphyrias
• HEP - Hepatoerythropoietic Porphyria
• Blisters
• EP - Erythropoietic Porphyria
• Facial hair
• HCP - Hereditary Coproporphyria
• Hyperpigmentation
• VP - Variegate Protoporphyria

PP AND AIP
• Porphyria: Defined as a group of rare inherited
PLUMBOPORPHYRIA
or acquired metabolic disorders caused by loss
ACUTE INTERMITTENT PORPHYRIA
or gain of function, and mutations in the enzyme
• Neurologic symptoms
responsible for the heme biosynthesis.
• Abdominal pain, vomiting, constipation and
o Deficiency or an excess can result in
tachycardia, and hypertension
Porphyrias
• SIMILARITY: Increase ALA and porphobilinogen
• Classifications of Porphyrias
(PBG) in urine
o Acquired or Inherited
o Black/Red in color urine that will turn
• Manifestations of Porphyrias (Another type of
into dark brown upon oxidation
classification)
▪ Color is due to porphyrin (red
o Neurologic symptoms
color urine) present or
o Cutaneous symptoms
porphobilin (black colored
o Or both
urine).
• Acquired porphyria
o Upon exposure to air or light it will turn
o PP - Plumboporphyria
into black or brown, from red.
o PCT - Porphyria Cutanea Tarda
• To differentiate, look for enzyme deficiency
• Inherited porphyria
o AIP - Acute intermittent Porphyria
PLUMBOPORPHYRIA (PP)
o CEP - Congenital Erythropoietic
Porphyria • Also known as ABP or ALA
o HEP – Hepatoerythropoietic Porphyria o Alpha-amino-lebolinic acid
o EP - Erythropoietic Porphyria dehydratase deficiency porphyria
o HCP - Hereditary Coproporphyria • ALA dehydratase-enzyme deficiency
o VP - Variegate Protoporphyria • Increased urine ALA, with a normal
• Porphyria that manifests neurologic symptoms porphobilinogen excretion.
o PP - Plumboporphyria • Can be mistaken as lead poisoning
o AIP - Acute intermittent Porphyria o Because lead poisoning also has an
• Porphyria that manifests cutaneous symptoms increased ALA present in the urine
o CEP - Congenital Erythropoietic o Most common cause of low ALAD
Porphyria activity is lead poisoning.
o PCT - Porphyria Cutanea Tarda • Differentiate PP and Lead poisoning
o HEP – Hepatoerythropoietic Porphyria o Add dithiothreitol or sulfhydryl reagent
o EP - Erythropoietic Porphyria when testing
• Porphyria that manifests both symptoms ▪ Lead poisoning has a normal
o HCP - Hereditary Coproporphyria ALAD upon usage of these
o VP - Variegate Protoporphyria reagents
• Example: from
• If a certain porphyria has a known increase in increased ALA, upon
ALA and porphobilinogen it will produce a addition of reagent, it
neuropsychiatric syndrome. returns to normal value
• If a certain porphyria has a known increase in ▪ PP will have no change at all at
neuro, copro, and the protoporphyrin it will ALA level
produce cutaneous symptoms. • Example is at first there
is increased ALA, then
NEUROLOGIC SYMPTOMS you added reagent
and still increased with
• Abdominal pain
same value, it means
• Vomiting
that the case is PP
• Constipation
• Tachycardia
Acute intermittent Porphyria (AIP)
• Hypertension
o Etc • Hydroxymethylbilane synthase (HMBS)
→ Related to neurons or the brain deficiency, also known as porphobilinogen
(PBG) deaminase.
 CONGENITAL ERYTHROPOIETIC PORPHYRIA (CEP) • Presence of scarring of the hands and the face
same thing as PCT maybe because of free
• Deficient: Uroporphyrinogen III cosynthase
radicals to either air, oxygen, or sunlight resulting
• Gunther’s disease
to oxidative damage
• URO and COPRO
• INCREASE ISOCOPRO and UROporphyrin
o Gunther’s Disease: There is a known
• Increase erythrocyte zinc protoporphyrin (ZPP)
increase in the URO and
COPROporphyrin present in urine
and/or sometimes the stool of the ERYTHROPOIETIC PROTOPORPHYRIA (EP)
patient.
• Erythropoietic Protoporphyria
o Erythrodontia: Teeth fluoresce red in UV
• Also presenting cutaneous symptoms
light) due to the deposition of an
o Photosensitivity
increased porphyrin in dentin which is
o Facial hair
present in the teeth.
o Burning, itching, pain in the skin upon
• Rarest and the most severe form of porphyria
exposure to light
• Manifestations can be seen as early as infants.
• Deficient: Ferrochelatase
o Urine of infants are red in color that turns
o The last enzyme involves in the heme
brown in color when oxidized, that is
synthesis pathway
observed in diapers
• Second most common porphyria
o Present cutaneous symptoms
• Burning of light exposed skin
▪ Cutaneous photosensitivity
• Increase FEP and Zn PROTO in RBC, plasma, and
among infants
stool
PORPHYRIA CUTANEA TARDA (PCT) • Hair growth serves as its defense mechanism
o There’s an excess growth of hair
• Deficient: Uroporphyrinogen decarboxylase
because of defense mechanism
• Most common porphyria
because when skin is exposed to light,
• Cutaneous symptoms
there will be blisters
o Blisters, fragility in light-exposed skin
• Excitation of porphyrins in the blood is another
(specially the hands).
thing upon exposure to sun
• Abnormal growth or hair
o Converted into 3 state porphyrins that
• ISOCOPRO, URO
provides available site for free radicals
o Diagnostically, there’s an increase in
o As free radicals react, oxidative
the ISOCOPRO and UROporphyrin and
damage in the tissue, particularly in the
the specimen of our patient
skin happens and that can cause blister
• Liver damage is possible because of iron
formation
depletion
• There’s blister formation in cutaneous symptoms HEREDITARY COPROPORPHYRINOGEN (HCP)
because with the air, the oxygen, and the
• Deficient: Coproporphyrinogen oxidase
sunlight, the free radicals present in the skin
• Hallmark is increase COPRO in stool (in urine too)
causes oxidative damage to the skin. Oxidative
• Increase in ALA and PBG
damage result to the formation of blisters
o Same with neurologic symptoms
classification of porphyria known as PP
TWO TYPES OF PCT
and AIP
Type I o Common in Harderoporphyria (rare
• SIMILARITY: Deficient in Uroporphyrinogen erythropoietic form of HCP)
decarboxylase VARIEGATE PORPHYRIA (VP)
• DIFFERENCE: It is restricted to the liver
Type II • South African Porphyria
• SIMILARITY: Deficient in Uroporphyrinogen o Presenting both neurological and
decarboxylase cutaneous symptoms (same as HCP)
• DIFFERENCE: There’s a deficiency in the tissue • Deficient: Protoporphyrinogen oxidase
• Manifestation is the same as Acute Intermittent
Porphyria (AIP)
HEPATOERYTHROPOIETIC PORPHYRIA (HEP) • Hallmark increase PROTO and COPRO in stool
• Hepatoerythropoietic porphyria
o Liver is affected Notes:
• Deficient: Uroporphyrinogen decarboxylase • Most of the symptoms of porphyria is light
o There’s a reduce activity of this enzyme sensitive
by 90% or more OTHER CUTANEOUS SYMPTOMS
• Photosensitivity begins in childhood
o Also presents cutaneous symptoms • Porphyrinuria
• Hepatic disease on later stage o Urine color is red because of presence
• There’s an excess facial hair present of porphyrin
• Presence of skin blisters in the face and hands Watson-Schwartz
• Erythrodontia
• In W-S Test, extraction method is done
o The teeth fluorescence red because of
o For extraction method: Butanol or
porphyrin accumulation in the dentin
Chloroform as reagent
• Scarring
o PBG from Urobilinogen or indole can be
a source of interference (you have to
consider that there could be a false
positive when it comes to result
SECONDARY PORPHYRIAS
because of those interfering
• Also known as Porphyrinurias substances)
• Increase excretion of Urinary porphyrins • Positive= Red color remains
• Not a defect in the biochemical synthesis of o If the red color remains in the aqueous
heme but a result of either: Caused by: phase when we perform this test, that is
o Other disorder (there’s an considered as positive for the presence
accumulation of porphyrins) of PBG
o Toxins
o Drug interfering heme synthesis Hoesch Test
pathway
• Increase ALA with normal PBG • Principle: PBG forms red-orange color when
o This one has normal PBG to differentiate mixed with Ehrlich’s reagent (para-
with other porphyrias Dimethylaminobenzaldehyde or pdab)
o There is normal PBG because heme o Reagent used here do not react with
synthesis not the problem urobilinogen that is why it is used as a
confirmatory test to watson-schwartz
TREATMENT • Ehrlich’s reagent component: (para-
Dimethylaminobenzaldehyde or pdab)
• We treat the different kinds of porphyrias • Positive = Red-Orange color formation
depending on the symptoms presented • Detects the PBG and ALA present in the urine
• Cutaneous • The reagent used in here do not react with
o If there is photosensitivity, avoid sunlight urobilinogen that’s why the hoesch test is used
o Sunbath agents as a confirmatory test to Watson-Schwartz
o Oral beta carotene to correct o Porphobilinogen and Urobilinogen are
whatever is the symptoms presented by known to produce a positive result
the patient when it comes to Watson-Schwartz
• Decreased Heme Load
o Desferrioxamine will be given to the Note:
patient to chelate irons so that irons will • Both tests will just determine whether
not accumulate in the blood porphobilinogen is present or not in the sample.
o phlebotomy • Porphobilinogen and urobilinogen are known to
• Other Treatments: produce a positive result on Watson-Schwartz
o Hematin- known to limit synthesis of • If you want to quantitatively measure the PBG
porphyrins in cells in the bone marrow. and ALA present, you can do so by performing
Depending on the symptoms iron exchange.
presented:
▪ If cutaneous, most of the
there’s treatment to that PORPHYRINS (URO, COPRO, PROTO)
▪ If neurologic, there could be Specimen:
treatment depending on the
symptoms or none at all • 24-hour urine, Whole Blood, 1 g fecal sample
o Those 3 porphyrins naka depende kung
saan sila mataas doon sila
LABORATORY DETERMINATION: matatagpuan
• Specimen Considerations:
PORPHYRIAS o Porphyrins are known to be light
sensitive
PBG AND ALA o Specimens should be protected from
→ Methods: Both are only a screening test, they light
are only a qualitative and not quantitative, the o Collect using an amber bottle/
purpose is to know if PBG and ALA are present in container or you can wrap the
a given specimen or not specimen container to prevent
o Watson-Schwartz exposure from any type of light. Any
o Hoesch test type of light degrades those porphyrins
o Store at 4 °C for 48 hours or
 o -20 °C for weeks if you cannot perform
the test within the day
▪ BUT you must perform the test
ASAP, once you get the
specimen
o BEST Anticoagulant: EDTA
o Collected and be transported at 4 °C in
a dark environment

Methods:
UV lamp or woods lamp (to screen)
• Screen test for the presence of porphyrins
• To check if the specimen will fluoresce.
• If it fluoresces it is an indication that there’s
presence of porphyrins
• Positive = orange-red fluorescence (for uro,
copro, proto)
Quantitative Methods: (to confirm)
• Fluorometry
o Quantitative
• HPLC
• HP-TLC
o sometimes spectrophotometer can
also be used wherein excitation and
emission wavelength are of concern.

If you really want to measure porphyrin we need to use:

• Excitation wavelength: 400-405 nm
• Emission wavelength: 594-598 nm
• Measurement is based on enhanced
fluorescence in acidic solution using that UV
lamp or woods lamp