NATIONAL UNIVERSITY-MANILA

MTAP 2 – Clinical Chemistry 2

DESCALSO, GONZALES, MANUEL, SOMERANO

ELECTROLYTES
 Ions capable of carrying an electric charge. Classified based on the type of charge they carry.
 Anion: negatively charged or Cation: positively charged.

 Functions
of Electrolytes
1. For volume and osmotic regulation
2. For myocardial rhythm and contractility
3. Important cofactors in enzyme activation
4. For the regulation of ATPase ion pumps
such as sodium and potassium
5. For neuromuscular excitability includes
magnesium and ionize calcium
6. For the production and use of ATP from
glucose such as phosphate
7. Maintenance of acid-base balance
8. Replication of DNA and the translation of
mRNA

Introduction: Water and Electrolyte Balance

TOTAL BODY WATER
 Electrolytes are actually affected by the
water content because it depends on the
number of solute from intracellular space
to extracellular space by the water
content.
 Water is considered to be the universal
solvent.
 The water in the body may varies from
male and female water content in the
body because the fat deposition.
 Majority of water is in the extracellular space and 2/3 are found in the cell
 Example of Electrolytes: Na+, Mg2+, HPO42-, K+, Cl-, HCO3-, SO42-, Ca2+
  Sodium and Potassium are considered THE CHIEF POSITIVELY CHARGED
CONSTITUENTS (cations).

 Chloride and Bicarbonate are considered THE CHIEF NEGATIVELY CHARGED

CONSTITUENTS (anions).

OSMOLALITY
 Based on the number of dissolved particles in a solution. Kung gaano kadami yung solute na
present sa blood.
 Measures the total concentration of all of the ions and molecules present in serum or urine
 Sodium, glucose, and urea are major contributors to the total osmolality of serum.
 REFERENCE RANGE: 275 to 295 mOsm/Kg.

Remember:
 The major solute that contributes to serum osmolality is SODIUM.
 Osmolality could be calculated and there’s 2 formula for calculating
osmolality.

Example:
 A 40 year old woman suffering from vomiting and diarrhea had the following laboratory values:
Na+ 145 mmol/L, glucose 750 mg/dL, BUN 25 mg/dL

OSMOLAL GAP
 The difference between the calculated osmolality and the measured osmolality
 Elevation in the gap is usually due to factors other than Na+, glucose, or BUN
 Presence of ketones or alcohol in the plasma can elevate the osmolal gap. Or osmotically
active for example mannitol which may increase the level of osmolal gap. Basta may
osmotically active na solute tataas osmolal gap.
 Osmolality is important because it is the condition to which the hypothalamus responds.
 The osmolality is triggered and maintained by our osmoreceptors which found in brain
particularly hypothalamus and when our body feels that there are many solutes, may
hyperosmolality or mababa dito nat’trigger kung magrerelease siya ng ADH, kung mag
aactivate ng RAAS.
 Clinical Importance: If a calculated osmolality is elevated above the reference range, the
patient is suffering from dehydration.
REGULATION OF BLOOD VOLUME

SODIUM
 CHARACTERISTIC
o Major extracellular cation
o Solute that contributes most to total serum osmolality
o Blood levels of sodium are mainly controlled by aldosterone
o Whenever sodium enters the cell, water follows.

 FUNCTIONS OF SODIUM
o Regulates osmolarity and blood volume
CAUSES OF HYPERNATREMIA CAUSES OF HYPONATREMIA
Diabetes insipidus Diuretics, Potassium depletion
Osmotic dieresis Aldosterone deficiency, ketonuria
Loss of thirst Salt-losing nephropathy, vomiting
Insensible loss of water Diarrhea, excess fluid loss as with burns, excess
Gastrointestinal loss of hypotonic fluid sweating or trauma
Excess intake of sodium SIADH, excess water intake
Adrenal insufficiency
Reset osmotat
Acute or chronic renal failure
Nephrotic syndrome, hepatic cirrhosis,
Congestive heart failure
Pseudohyponatremia (hyperglycemia,
hyperlipidemia, hypernaprotenemia)
  SPECIMEN CONSIDERATIONS
o Serum, plasma, 24-hour urine sample, as well as sweat and CSF can be used as
sample
o If plasma is used, lithium heparin and ammonium salts of heparin, as well as lithium
oxalate may be used
o Heparin is the anticoagulant of choice specially for electrolyte determination
o Hemolysis does not cause significant changes but marked hemolysis should be avoided
o The equation can be used to check on accuracy of electrolyte determination:

Na = CO2 + Cl + 10 or Na = CO2 + Cl + 12
 METHODOLOGIES
o Flame Emission Photometry
 Sodium produces yellow color when exposed to flame. Pag nakakita ka ng
fireworks na yellow, sodium yon par.
 Sodium emits light at a wavelength of 590 nm
 Dilute sample first to prevent interferences to prevent atomizer plugging and
acquire increased sensitivity
 Dilute sample using high purity water (deionized water with electrolyte content of
<0.05 ppm)
 Serum is usually diluted 1:100 or 1:200
 Lithium or cesium may act as internal standard
o Atomic Absorption Spectrophotometry
o Ion-selective electrode (glass aluminum silicate) – membrane used
 Reference value:
o Serum: 135 to 145 mmol/L Conversion Factor: 1.0 (sodium, potassium, chloride and
lithium)

POTASSIUM
 Characteristic
o Major intracellular cation
o Chief counter-current of Sodium
o PISO – Potassium IN, Sodium OUT
 Functions of Potassium:
o Involved in proper transmission of nerve impulses
o Important for contraction of the heart –abnormal levels of potassium can lead to altered
electrocardiographic patterns. Whenever, there is Hyperkalemia it may cause an
abnormality in ECG, because potassium makes an important role in contraction.
 Clinical Considerations
Causes of Hyperkalemia Causes of Hypokalemia
Decreased renal excretion GI Loss
Acute or chronic renal failure Vomiting, diarrhea, gastric suction,
Hypoaldosteronism; Addison’s disease intestinal tumor, malabasorption
Diuretics Cancer therapy – chemotherapy, radiation
Increased intake Therapy
Oral or IV K replacement therapy Large doses of laxatiives
Cellular shift Decreased intake
Acidosis, Muscle/cellular injury Renal loss
Chemotherapy Diuretics-thiazides, mineralocorticoids
Leukemia (increased WBC) Nephritis, renal tubular acidosis
 Hemolysis Hyperaldosteronism; Cushing’s syndrome
Increased intake Hypomanesemia
Sample hemolysis, thrombocytosis Acute leukemia
Prolonged tourniquet application or Cellular shift
Excessive fist clenching Alkalosis, Insulin overdose

 Specimen Consideration
o Serum or plasma can be used; plasma/serum must be separated from the cells quickly to
prevent potassium from shifting from RBCs to serum
o Heparin is the anticoagulant of choice
o Whole blood samples for potassium determination should be stored at room temperature
o NO to prolonged tourniquet application, excessive fist clenching and hemolysis
o Pseudohyperkalemia – this are factors that may cause hyperkalemia because of improper
specimen collection. Major cause is hemolysis.
o The specimen should not place on ice. When place in ice, the platelet releases
Potassium. The sample should be in ROOM TEMPERATURE
 Methodologies
o Flame Emission Photometry
 Potassium produces violet color when exposed to flame
 Potassium emits light at a wavelength of 768 nm
 Dilute samples first to prevent interferences, to prevent atomizer plugging and to
acquire increased sensitivity
 Dilute sample using high purity water
 Serum is usually diluted 1:100 to 1:200
 Lithium or cesium may act as internal standard
 Atomic Absorption spectrophotometry
 Ion selective electrode (valinomycin membrane)
o Reference Value
 3.4 to 5.0 mmol/L Conversion Factor: 1.0

CHLORIDE
 Characteristic
o Major extracellular anion
o It is the chief counter ion of sodium in
ECF. Pag pumasok si sodium, sasabay
si chloride.
o It is the only anion to serve as an
enzyme activator. AMYLASE requires
calcium and chloride.
 Functions
o Maintains water balance, osmotic
pressure and anion-cation balance in the
extracellular fluid
o Responsible for chloride shift – an
exchange mechanism between chloride and bicarbonate across the membrane of
RBCs.
  Clinical Considerations
HYPERCHOLEREMIA HYPOCHOLEREMIA
Excess loss of bicarbonate Prolonged vomiting, Diabetic ketoacidosis,
Renal tubular acidosis aldosterone deficiency
Metabolic acidosis Salt-losing renal diseases (pyelonephritis)
High serum bicarbonate (compensated
respiratory acidosis or metabolic alkalosis)

 Specimen Consideration and Patient Preparation:

o Serum, plasma, whole blood, 24 hour urine and sweat can be used as sample. Disorder
that is characterized by extreme production of electrolytes in sweat using a patch test is
called or indicative of CYSTIC FIBROSIS particularly the chloride.
o Lithium heparin is the anticoagulant of choice
o Marked hemolysis should be avoided
 Methodologies
o Ion selective electrode
 Most commonly used; using an ion exchange membrane selective for chloride
ions. Membrane used is a combination of silver wire coated with AgCl.

o Amperometric-Coulometric Titration
 Principle of Cotlove chloridometer
 Uses coulometric generation of silver ions which combine with chloride to
quantitate chloride concentration
 Excess silver ions, which were not bound to chloride is used to indicate endpoint.
 Sample diluted in acid with small amount of gelatin
 Nitric acid provides good conductivity
 Acetic acid provides sharper endpoint by reducing solubility of silver
chloride by decreasing polarity.
 Gelatin makes a smoother titration curve by equalizing the reaction rate
over the entire electrode

o Mercuric Titration
 Principle of S chales and Schales
 Based on the reaction of chloride ions to mercuric ion s to form mercuric chloride
 Blood containing bromide leads to positive error.
 Which of the following interferences causes false positive result on
chloride determination? Ans: BROMIDE in Schales and Schales causes
positive error. It falsely increases the level of chloride
 Excess mercuric ions are then made to react with diphenylcarbazone in order to
form violet blue color which is endpoint of Schales and Schales.

o Colorimetric Method
 Uses mercuric thiocyanate and ferric nitrate to form ferric thiocyanate, which
is reddish colored complex with a peak absorbance at 480 nm.
 Used in autoanalyzer (Technicon)
o Reference Values
 Serum: 98 to 107 mmol/L Conversion factor: 1.0
CALCIUM
 Characteristic
o 99% of calcium is found in bones and teeth; 1% is found in the blood
o Calcium exist in three forms:

o 1 g/dL decrease in albuim causes 0.8 mg/dL decreased in total calcium

o Calcium is absorbed in ileum at acid pH in the presence of Vit. D
 Function of Calcium
o Contributor to structure of bone and teeth
o Coagulation (calcium is coagulation factor IV)
o For proper contraction of heart and muscles
o Activator to enzymes
o Neurotransmission regulator
 Clinical Considerations

 Specimen Considerations
o Serum, plasma or 24-hour urine sample maybe used; Lithium heparin is preferred
o No to EDTA, oxalate and citrate because calcium is the specimen
o Samples should be collected anaerobically
o For 24-hour urine calcium, sample should be acidified using 6M HCl (1 mL of HCl per
100 mL of urine)
o For assay
 Methodologies
o Orthocresolphtalein complexone
 A calcium chelator; produces reddish complex with absorbance at 570-578 nm
 8-hydroquinoline binds magnesium which may interfere
 Urea can be used to decrease the turbidity of lipemic serum and increase
intensity of the calcium dye complex
 Ethanol can be used to decrease the absorbance of the blank
o Arsenazo III
o Alizarin
o Methylthymol blue
 Atomic Absorption Spectrophotometer
o Calcium compounds in a flame dissociate into free calcium atoms
o Free atoms absorb light of a characteristic wavelength
o Lanthanum is used to bind phosphate that might instead bind the calcium and cause
falsely low result
  Ion selective electrode
 Clark Collip Precipitation Method
o Classic method that measures oxalic acid as the end product
 Ferro Ham Chloroanilic acid Precipitation Method
o Precipitation of calcium with chloroanilic acid
 Reference Value
o Total Calcium (child) = 2.20-7.50 mmol/L (adult)= 2.15-2.50 mmol/L
o Ionized Calcium (child) = 1.20-1.38 mmol/L (adult) = 1.16-1.32 mmol/L
 Conversion factor = mg/dL to mmol/L 0.25

MAGNESIUM
 Characteristic
o Second most abundant intracellular cation (after potassium)
o Fourth most abundant cation in the body
o 50% of magnesium is found in the bone; 25% is in the muscle
o Exists in the blood in three forms:
 Free magnesium = 55%
 Complexed magnesium = 15%
 Protein bound = 30%
 Functions of Magnesium
o Contributor to bone structure
o For muscle contraction and heart rhythm
o Activator to enzymatic reactions
o During tetani, if the level of ionized calcium may cause involuntary muscle movement
called tetani. Moreover, hypomagnesemia may cause tetani.

 Clinical Considerations

 Specimen Consideration
o Serum, lithium heparinized plasma or 24-hour urine may be used
o Oxalate, EDTA and citrate should not be used
o No to hemolysis
o 24-hour urine should be acidified with HCl to prevent precipitation
  Methodologies
o Colorimetric Method
 Calmagite (Hitachi and Synchron)
 A naphtol sulfonic acid derivative
 Use of polyvinylpyrrolidone minimizes the effects of serum protein
 Mg+ calgamite = reddish violet complex read at 520-532 nm
 Strontium chelate = masks the effect of calcium
 Triethanolamine = to mask the effect of iron
 Formazan dye (vitros) – colored complex at 660 nm
 Methyl-thymol blue (Dimension, DuPont aca)
 Dye-lake method – Titan yellow dye (Clayton yellow/thiazole yellow); titan yellow
forms a red lake with magnesium; polyvinyl alcohol increases the sensitivity of
the method
 Fluorometry – magnesium reacts with the reagent 8-hydroxy-5-quinoline
sulfonic acid or calcein to form a fluorescent compound (390-410 nm)
 AAS – reference method
 Ion-selective electrode
 Reference Value
 0.63-1.0 mmol/L
 Conversion factor = mEq/L to mmol/L 0.5

PHOSPHATE
 CHARACTERISTIC
o Intracellular anion; most phosphorus is in the form of phosphate
o Most serum phosphate are inorganic; most phosphorus inside the cell is inorganic
o Phosphate metabolism is controlled by parathyroid hormone, calcitonin and Vitamin
D
o Exists in the blood in three forms
 Free phosphate = 55%
 Complexed phosphate= 35%
 Protein bound= 10%
 Functions of Phosphate
o Serves as a buffer (biphosphate-dihydrogen phosphate buffer system) 4:1 to maintain
the pH na 7.35 at 7.45
o Serves as a part of energy molecules like ATP
 Clinical Consideration

 Specimen Consideration
o Serum, lithium heparin plasma or 24-hour urine sample may be used
o Oxalate, EDTA and citrate should not be used
o No to Hemolysis
o Diurnal variation = highest levels are found in late morning; lowest in the evening
  Methodologies
o Fiske-Subbarow Method
 Uses molybdate reagent; products that can be measured include:
 Measurement of ammonium molybdate complex at 340 nm
 Reduction to form molybdenum blue, which is read at 660 nm; reducing agents
that can be used in this reduction process include: ANSA, stannous chloride,
ascorbic acid and N-phenyl-phenylenediamine
 Serum proteins are precipitated by trichloroacetic acid and phosphate is converted into
phosphomolybdate comple (MoVI) by the addition of sodium molybdate . The addition of p-
methylaminophenol reduces the (MoVI) into (MoV). The absorbance of the solution at 700 nm is
proportional to the serum phosphate concentration.

 Reference Value
o Serum/Plasma
 (neonate) 1.45-2.91 mmol/L
 (child) 1.45-1.78 mmol/L
 (adult) 0.87-1.45 mmol/L
 Conversion Factor
o mg/dL to mmol/L = 0.323

ANION GAP
 Difference between unmeasured anions and unmeasured cations
 Useful in indicating an increase in one or more of the unmeasured anions in the serum
 Serves as a form of quality control for the analyzer used to measure these electrolytes
 Formula
o AG = Na – (Cl + HCO3) or AG = (Na + K) – (Cl + HCO3)
 Reference Value
o 7 to 16 mmol/L or 10 to 20 mmol/L
 Clinical Significance
o Increased AG - uremia/renal failure, ketoacidosis, poisoning due to ingestion of toxic
substances like methanol, ethanol, ethylene glycol poisoning or salicylate; lactic
acidosis; severed dehydration; instrument error
 MUDILES – common causes of increased anion gap
o Methanaol
o Uremia
o Diabetic ketoacidosis
o Iron/inhalants (i.e. carbon monoxide, cyanide, toluene), isoniazid, ibuprofen
o Lactic acidosis
o Ethylene glycol poisoning , ethanol ketoacidosis
o Salicylates, starvation ketoacidosis, sympathomimetics
o Decreased AG – rare, hypoalbuminemia (decreased in unmeasured anions); severe
hypercalcemia (increase in unmeasured cations); patients with multiple myeloma;
instrument
ACID BASE – BLOOD GASES
Definitions
 Arrhenius’ Definition
o An acid is a substance that increases the concentration hydrogen ion (H +) when dissolved
in water
o A base is a substance that increases the concentration of hydroxyl ion (OH-) when
dissolved in water
 Bronsted and Lowry’s Definition
o An acid is a substance that donates a proton in a reaction
o A base is a molecule that donates a pair of electrons for a covalent bond
 Lewis’ Definition
o An acid is a molecule or ion that accepts a pair of electrons to form a covalent bond
o A base is a molecule that donates a pair of electrons for a covalent bond
 Dissociation Constant
o Also known as ionization constant K value, describes the relative strengths of acids and
bases
 pK
o Negative log of ionization constant and pH in which protonated and unprotonated forms are
present in equal concentrations

ACID-BASE BALANCE
 All the chemical and metabolic reactions are pH dependent, hence an alteration in acid-
base status can lead to alterations in consciousness, neuromuscular irritability, tetany, coma
and death
 Important in order to maintain the pH within the normal range (normal range of pH = 7.35 –
7.45)
 Important to maintain homeostasis, important in enzyme function
 Maintenance of H+
o Normal concentration of H in extracellular body fluid ranges from 36 to 44 nmol/L, but
body produces much greater quantities of H
o Via lungs and kidneys, body controls and excretes H to maintain pH homeostasis
o Acidosis: a pH level below reference range (<7.34)
o Alkalosis: a pH level above reference range (>7.45)

BUFFER SYSTEM
 Buffers are substances that resist change(s) in pH
 A buffer system is composed of a “weak base and its conjugate acid” or a “weak acid and its
conjugate base”
 Buffer System in the Body
o Bicatbonate-carbonic acid buffer system
 The most important buffer system in the body
 Bicarbonate-carbonic acid ratio must be 20:1 in order to maintain normal pH
 Over 90% of blood carbon dioxide exists in the form of bicarbonate ion
o Biphosphate-dihydrogen phosphate buffer system
 Must be maintained at a ratio of 4:1
o Hemoglobin - since it transports gases (oxygen and carbon dioxide)
 o Plasma proteins – since proteins are amphoteric (have negative and positive charge)
 Buffer systems are body’s first line of defense against extreme changes in H concentration
 All buffers consist of a week acid & its salt or conjugate base.
 Bicarbonate-carbonic acid system has low buffering capacity, but is still important buffer for 3
reasonss:
o H2CO3 dissociates into CO2 and H2O, allowing CO2 to be eliminated by lungs and H as
water
o Changres in CO2 modify ventilation (respiration) rate
o HCO3- concentration can be altered by kidneys
o Other buffers: phosphate system & plasma protein
o
REGULATION OF ACID-BASE BALANCE
 Lungs
o Lungs help maintain acid-base balance through gas exchange or respiration
o Rapid and short term compensation either hypoventilate andhyperventilate,
respectively.
o Analyte(s) controlled: O2 and CO2

 Interrelationship of bicarbonate & hemoglobin buffering systems

  In hypoventilation, CO2 is not eliminated at the rate of its production causing an increase in H+
concentration
 If CO2 removal faster than usual (hyperventilation), the H+ will decrease
 Consequently, ventilation affects the pH of the blood
 Alterations in H+ concentration due to non-respiratory insults causes the respiratory center to
respond by altering the rate of ventilation
 The lungs compensate immediately (first line of defense to changes in acid-base status) but
the response is SHORT-TERM and INCOMPLETE

 Kidneys
o Kidneys help maintain acid-base balance through reabsorption or excretion of
bicarbonate.
o Slow but long term compensation and complete; analyte controlled: bicarbonate (HCO3-
)
1. Reclamation of bicarbonate (almost 100%) in the PCT in the form of CO2 (most important role
2. Excretion of acids in the form of:
a. Ammonium ions (NH4+) (2/3)
b. Dihydrogen phosphate (H2PO4-)
c. Titratable acids (free H+)
3. ATP-dependent excretion of H+ in exchange of sodium via Na+ - H+ pumps
 The kidneys are slower to respond (2-4 days), however the response is long-term and
potentially complete.

OXYGEN-HEMOGLOBIN DISSOCIATION CURVE

 O2 must be released at tissues from its carrier, hemoglobin.

 Oxygen dissociates from adult (A1) hemoglobin in characteristic fashion (S-shaped curve)
 Shape of oxygen-
dissociation curve & affinity
of hemoglobin for O2 are
affected by:
o Hydrogen ion
activity
o PCO2 & CO levels
o Body temperature
o 2,3-DPG
HENDERSON-HASSELBACH EQUATION
 States that relationship between lungs, kidneys and pH
 Bicarbonate = total carbon dioxide minus Carbonic acid
 Carbonic acid = partial pressure of carbon dioxide x 0.031
 Formula:

 pH is directly proportional to bicarbonate – an increase in bicarbonate causes and increase in

pH and vice versa
 pH is indirectly (inversely) proportional to pressure of carbon dioxide- an increase in pCO2
causes a decrease in pH and vice versa

CLINICAL SIGNIFICANCE
 Acidosis refers to a decrease in blood pH
 Alkalosis refers to an increase in blood pH
 Changes in pH can be causes by either defect in the lungs (respiratory) or defect in the
kidneys (metabolic)
 When one organ has a problem, the other organ will compensate. That means when lungs
have problem, the kidneys will compensate. When kidneys have problem, the lungs will
compensate
DETERMINATION OF ACID-BASE STATUS
1. Evaluate the pH
 Normal pH: 7.35 to 7.45
o < 7.35 = Acidemia
o > 7.45 = Alkalemia
 pH 7.4 = optimum value for arterial blood
 An increase in H+ concentration decreases pH, whereas a decrease in H+ concentration
increases pH/
 The pH decreases by 0.015 each Celsius above 37 C
2. Evaluate the ventilation (Lungs) pCO2
 Normal pCO2: 35-45 mmHg
o < 35 mmHg = Respiratory alkalosis
o > 45 mmHg = Respiratory acidosis
 The lungs regulate pH through the retention or elimination of CO2
 Barbiturates, morphine or alcohol increase pCO2
 An increasing ratio of hepatin to blood can cause artifactual rise on measured pCO2
(12-15%)
o For ABG, 0.05 mL of liquid heparin (1000 IU/mL) should be used for each milliliter of
blood
3. Evaluate the metabolic process (Kidneys)
HCO3-
 Normal HCO3: 21-28 mEq/L
o < 21 mEq/L = Metabolic acidosis
o > 28 mEq/L = Metabolic alkalosis
4. Evaluate the degree of oxygenation
 Normal pO2 = 80-110 mmHg (adequate oxygenation
 Low pO2 = Hypoxemia
o Mild hypoxemia = 61-80 mmHg
o Moderate hypoxemia = 41-60 mmHg
o Severe hypoxemia = 4o mmHg
 For pO2 values between 70 and 100 mmHg, the saturation of hemoglobin is close to 100%
Parameters useful in assessing oxygen status
1. Oxygen saturation (SO2)
2. Fractional (percent)
oxyhemoglobin (FO2Hb)
3. Trends in oxygen saturation using
transcutaneous pulse oxymetry
(SpO2)
4. pO2
Specimen Collection
 Specimen: arterial whole blood using heparin as anticoagulant
 Venous blood is usually 0.03 pH units lower than arterial blood
 Arterialized venous blood may be obtained by heating the hand and forearm in water at 45 C
for 5 minutes and then drawing blood from the dilated veins on the back of the hand
 Capillary blood is arterialized by warming the ear, finger, or heel at 45 C before taking the
sample
 Syringe with rubber stopper; specimen should be sealed
 Anaerobic collection
 Do not use vacutainer tube
 Place specimen in ice water or ice bath
 No to clots, no to hemolysis, no to bubbles
 Measurements are done at 37±0.05 C (Kaplan)
 For each degree of fever in the patient, pO2 will fall 7% and pCO2 will rise 3%
 If pH and blood gases are to be done within 20 minutes, no refrigeration is necessary

CASES RELATED TO BLOOD GAS ANALYSIS

 Specimen was exposed to room air
o Increase in oxygen, decrease in carbon dioxide, increase in pH
o Room air is composed of 20-22% oxygen. This atmospheric air may enter the specimen
causing an increase in oxygen while displacing carbon dioxide in the process – leading
to a decrease in carbon dioxide in the specimen.
 Sealed specimen was left at room temperature
o Decrease in oxygen, increase in carbon dioxide, decrease in pH
o Changes are due to the presence of blood cells utilizing glucose and oxygen at room
temperature, causing the formation of acid products and carbon dioxide
 Excess Heparin
o Heparin is an acid mucopolysaccharide
o It is often used at a concentration of 0.2 mg/mL of blood
o Excess of acid mucopolysaccharide leads to acidic pH of blood specimen

Methodology
 pH – glass electrode connected to a reference electrode (calomel electrode, mercury-
mercuric chloride)
 pCO2 – Severinghaus electrode – A modified pH electrode; glass electrode with weak
bicarbonate solution enclosed in silicone membrane
 pO2 – amperometric/polarographic; Clark electrode – composed of oxygen permeable
membrane (i.e., Teflon, polyethylene) with electrode composed of a platinum cathode and
silver-silver chloride anode
 Bicarbonate and Carbon Dioxide content may be obtained by nomogram from blood gas
analyzers
 CO2 content – consists of bicarbonate, undissociated carbonic acid, dissolve carbon dioxide
and carbamino-bound carbon dioxide
 Methods for CO2 content
o Automated Enzymatic Method
 All forms of CO2 are converted to bicarbonate by addition of base
 Bicarbonate is converted to oxaloacetic acid using phosphoenol pyruvate
carboxylase
 Acid malate dehydrogenase and measure consumption of NADH at 340 nm, as
oxaloacetic acid is converted to malate
o Automated Colorimetric Method
 Bicarbonate, carbonic acid and carbamino-bound carbon dioxide is released by
addition of acid
 Gaseous carbon dioxide is dialyzed through a silicone-rubber gas-dialysis
membrane into a buffer solution of cresol-red at pH 9.2
 Decrease in color intensity is proportional to the carbon dioxide content
 Decrease in color intensity is measured at 430 nm
 Other continuous flow methods uses phenolphthalein as indicator.
o Gasometric (Van Slyke or Kopp-Natelson)
 Determines the amount of physically dissolved carbon dioxide and amount of
carbon dioxide released from bicarbonate and carbonic acid
 Gaseous carbon dioxide present can be measured using:
 Volumetrically – volume of gas at atmospheric pressure
 Manometrically – pressure of gas at a fixed volume
 Reagents:
o Lactic acid = releases carbon dioxide
o Caprylic alcohol = prevents foaming
o Sodium hydroxide = absorbs carbon dioxide
o Autoanalyzer (Autotechnicon)
 Reagents:
 Sulfuric acid = releases carbon dioxide
 Buffered phenolphthalein = absorbs carbon dioxide
o Oxygen Content and Percent Oxygen Saturation
 Methods for Oxygen Content
 Spectrophotometric – measurement of absorbance of a hemolyzed
blood sample at two wavelengths (650 and 805 nm)
 Oximetric – uses Pulse Oximetry; noninvasive; light shines through a
finger or the bridge of the nose to a detector and absorbance is measured
at 650 and 805 nm
 Gasometric – manometric method devised by Van Slyke
o Transcutaneous pO2 and pCO2 – for continuous monitoring of partial pressures of
oxygen and carbon dioxide on a noninvasive basis
VITAMINS AND TUMOR MARKERS WITH TRACE ELEMENTS
 VITAMINS
o Are essential organic substances that are required in microgram to milligram amounts for
health, growth and reproduction
o Functions: antioxidants, enzyme cofactors, hormones and important in blood cell
maturation, bone formation and active in energy metabolism
TRACE ELEMENTS
 Are metals, except for selenium and the halogens, fluoride and iodine
 They are in tissue concentrations of less than 1 ug/g of wet tissue and constitute less than 0.01%
of dry body weight.
 Classified as essential and non-essential, yung non-essential they are usually toxic in the body.
Essential elements, whereas, there’s a deficiency has been supplied by a corresponding element
and corrects the deficiency.
TUMOR MARKERS
 Are substances in the body that is associates with the presence of cancer.
 There is no specific tumor marker
 There is no screening test that is use for tumor marker except for Prostate Specific
Antigen. The rest are usually use for monitoring and recurrence of cancer
 It may be enzymes, hormones, receptors, oncofetal antigens or oncogenes
 Analysis is mostly essential to test recurrence of cancer.

HCG – for males: testicular

carcinoma
THERAPEUTIC DRUG MONITORING
 Anticonvulsants are also known as anti-epileptic drugs
 Phenobarbital is used in the treatment of generalized grand-mal tonic-clonic seizures
(Henry’s 23rd ed). Is that both sides of the brain, there is an increase electric current
 Lithium salt (lithium carbonate) is used for the treatment of bipolar disease or manic
depression
 Bronchodilators are also known as anti-asthma drugs. Theophylline is an anti-asthmatic
drug that is teratogenic. Its characteristic is 50% protein bound
 Chloramphenicol is often associated with aplastic anemia and causes blood dyscrasia
 Aspririn (acetylsalicylic acid) is an analgesic, anti-inflammatory and anti-pyretic drug
 Salicylate toxicitiy can be checked using a colorimetric method utilizing Trinder’s reagent
which is composed of mercuric chloride, hydrochloric acid and ferric nitrate. Mercuric salt
precipitates proteins present in specimen. Ferric ions form purple complex with salicylate
derivatives which can be measured at 540 nm.
 Acetaminophen can be detected using nitrous acid (sodium nitrate + hydrochloric acid).
Addition of the reagent causes formation of 2-nitro-5-acetaminophenol which turns into a
yellow color in alkali. The yellow color is measured at 430 nm.

STAGES OF ETHANOL IMPAIRMENT (KABISADUHIN. LUMALABAS SA BOARD EXAM)