Enzymes

Enzyme concentration
Substrate
concentration
Saturation kinetics
Cofactors
Coenzymes
Activators

Metalloenzymes
Inhibitors
Competitive inhibitor
Noncompetitive
inhibitor
Uncompetitive inhibitor
Isoenzymes
Temperature
40-50C
60-65C
Temperature coefficient
(Q10)
pH
Storage
Hemolysis
Lactescence or milky
specimen
Enzyme nomenclature
Enzyme classification

Oxidoreductases

Serum
Enzyme concentration = reaction rate
Reagent
If enzyme > substrate, substrate = reaction rate
When substrate concentration reaches a maximal value, higher
concentration of substrate no longer results in increased rate of
reaction
Nonprotein entities
Organic compound
Ex. NADP
Coenzyme = Velocity
Inorganic ions
Alters spatial configuration of the enzyme for proper substrate
binding
Ex. Ca2+ (#1 activator), Zn2+ (LDH), Cl- (AMS), Mg2+ (CK, ALP)
Inorganic ion attached to a molecule
Ex. Catalase, cytochrome oxidase
Interferes with the enzymatic reactions
Binds to the active site of an enzyme
Reversible (Substrate > Inhibitor)
Binds to the allosteric site (cofactor site)
Irreversible
Binds to the enzyme-substrate complex
Substrate = ES = Inhibition
Same catalytic reactions but slightly different molecular structures
Fractionation of isoenzymes
37C = optimum temperature for enzyme activity
Temperature = Reaction rate (movement of molecules)
Denaturation of enzymes
Inactivation of enzymes
For every 10OC increase in temperature, there will be a two-fold
increase in enzyme activity
Most physiologic reactions occur in the pH range of 7-8
Enzymes: -20C = for longer period of time
Substrate and Coenzymes: 2-8C
LDH (LD4 & 5): Room temperature
Mostly increases enzyme concentration
Decreases enzyme concentration
1st digit: classification
2nd and 3rd digits: subclass
4th digit(s): serial number
OTHLIL
Oxidoreductases
Transferases
Hydrolases
Lyases
Isomerases
Ligases
Redox reaction
Dehydrogenases:
lec.mt 04 |Page | 1

Transferases

Hydrolases

Lyases

Isomerases
Ligases
Active site
Allosteric site
Prosthetic group
Holoenzyme
Zymogen/proenzyme
Emil Fishers/Lock and
Key theory
Kochlands/Induced fit
theory
Enzyme kinetics
Absolute specificity
Group specificity

-Cytochrome oxidase
-LDH
-MDH
-Isocitrate dehydrogenase
-G-6-PD
Transfer of a chemical group other than hydrogen from 1 substrate
to another
Kinases, Transaminases, Aminotransferases:
-CK
-GGT
-AST
-ALT
-OCT
Hydrolysis/splitting by addition of water
Esterases:
-ACP
-ALP
-CHS
-LPS
Peptidases:
-Trypsin
-Pepsin
-LAP
Glycosidases:
-AMS
-Galactosidases
Removal of groups w/o hydrolysis (product contains double bonds)
Aldolase
Decarboxylases:
-Glutamate decarboxylase
-Pyruvate decarboxylase
-Tryptophan decarboxylase
Intramolecular arrangements
Glucose phosphate isomerase
Ribose phosphate isomerase
Joining of 2 substrate molecules
Synthases
Water-free cavity
Where the substrate interacts
Cavity other than the active site
May bind regulatory molecules
Coenzyme that is bound tightly to the enzyme
Apoenzyme + Prosthetic group
Inactive form of enzyme
Shape of the key (substrate) must fit into the lock (enzyme)
Based on the substrate binding to the active site of the enzyme
Acceptable theory
Enzymes catalyze reactions by lowering the activation energy level
that the substrate must reach for the reaction to occur
Enzyme combines w/ only 1 substrate and catalyzes only 1
reaction
Enzymes combine w/ all the substrates in a chemical group
lec.mt 04 |Page | 2

Bond specificity
Zero-order reaction
First-order reaction
Measurement of
enzyme activity
International Unit
Katal Unit
Nonkinetic assay
Alkaline Phosphatase

Phenylalanine
L-leucine
Levamisole
3M urea
Methods (ALP)

Increased ALP
Acid Phosphatase
Prostatic ACP
RBC ACP
Methods (ACP)

Aspartate
Aminotransferase
(AST/SGOT)

Enzymes reacting w/ specific chemical bonds

Reaction rate depends only on enzyme concentration
Independent on substrate concentration
Reaction rate is directly proportional to substrate concentration
Independent on enzyme concentration
Change in substrate concentration
Change in product concentration
Change in coenzyme concentration
1 micromole of substrate/minute
1 mole of substrate/second
Absorbance is made at 10-second intervals for 100 seconds
pH = 10.5
405nm
Electrophoresis:
(+) Liver Bone (Regan) Placenta Intestine (-)
Heat fractionation:
( Stable) Regan Placenta Intestine Liver Bone ( Labile)
Inhibits Regan, placental and intestinal ALP
Inhibits Nagao ALP
Inhibits liver and bone ALP
Inhibits bone ALP
Low temperature = Increased ALP
1. Bowers and McComb (PNPP) IFCC recommended
2. Bessy, Lowry and Brock (PNPP)
3. Bodansky, Shinowara, Jones, Reinhart = BGP (beta
glycerophosphate)
4. King and Armstrong = PP (phenylphosphate)
5. Klein, Babson & Read = Buffered PPP (phenolphthalein
phosphate)
6. Huggins and Talalay = PPDP (phenolphthalein diphosphate)
7. Moss = ANP (alpha naphthol phosphate)
Sprue
Hyperparathyroidism
Rickets (children) and osteomalacia (adults)
pH = 5.5
405nm
Sources: Prostate (major), RBC, platelets, bone
Inhibited by L-tartrate ions
Inhibited by cupric and formaldehyde ions
Room temperature (1-2 hrs) = decreased ACP
Thymolphthalein monophosphate = specific substrate, substrate of
choice (endpoint)
Alpha-naphthyl phosphate = preferred for continuous monitoring
methods
1. Gutman and Gutman = PP
2. Shinowara = PNPP
3. Babsonm Read and Phillips = ANP (continuous monitoring)
4. Roy and Hillman = Thymolphthalein monophosphate (endpoint)
pH 7.5
340nm
Sources: Cardiac tissue > Liver > Skeletal muscle > Kidney,
pancreas, RBCs
lec.mt 04 |Page | 3

Alanine
Aminotransferase
(ALT/SGPT)
Methods (AST and ALT)

Increased
Transaminases

Amylase

Methods (AMS)

Saccharogenic
Amyloclastic
Chromogenic
Coupled-enzyme
Lipase
Methods (LPS)

Lactate dehydrogenase

Methods (LDH)

10-fold increase (LDH)

2-3x URL
Creatine Kinase

Duchennes muscular

pH 7.5
340nm
Major Source: Liver
1. Karmen method = Kinetic
2. Reitman and Frankel = Endpoint
-Color developer: DNPH
-Color intensifier: 0.4N NaOH
DeRitis ratio (ALT:AST) >1.0 = Acute hepatitis (Highest)
20x = viral or toxic hepatitis
Moderate elevation = chronic hepatitis, hepatic cancer, IM
Slight elevation = Hepatic cirrhosis, alcoholic hepatitis, obstructive
jaundice
Smallest enzyme (appears in urine)
Earliest pancreatic marker
P3: most predominant pancreatic AMS isoenzyme in AP
Isoenzymes:
S-type (ptyalin): anodal
P-type (amylopsin): cathodal
Samples w/ high activity of AMS should be diluted w/ NaCl to prev.
inactivation
Salivary AMS = inhibited by wheat germ lectin
Substrate: Starch
Reducing sugars produced
Classic reference method (SU)
Degradation of starch
Increase in color intensity
Continuous-monitoring technique
Late marker (AP)
Most specific pancreatic marker
Substrate: Olive oil/Triolein
1. Cherry Crandal (Reference method)
2. Tietz and Fiereck
3. Peroxidase coupling (most commonly used method)
Lacks specificity
RBC: 150x LDH than in serum
Sources:
LD1 (-HBD) and LD2 = Heart, RBC, Kidneys
LD3 = pancreas, lungs, spleen
LD4 an LD5 = liver and muscle
LD6 = alcohol dehydrogenase
1. Wacker method (forward/direct) = pH 8.8, 340 nm, most
commonly used
2. Wrobleuski LaDue (reverse/indirect) = pH 7.2, 2x faster
3. Wrobleuski Cabaud
4. Berger Broida
Hepatic carcinoma and toxic hepatitis
Viral hepatitis and cirrhosis
Isoenzymes:
CK-BB = most anodal, brain
CK-MB = myocardium (20%)
CK-MM = least anodal, skeletal and smooth muscles (Major, 94100%)
Total CK: 50x URL (highest)
lec.mt 04 |Page | 4

dystrophy
CK-MB
Methods (CK)
Adenylate kinase
N-acetylcysteine
Liver cells and RBC
Clelands reagent and
glutathione
Electrophoresis
CK relative index (CKI)
Aldolase

5 Nucleotidase

GGT

Methods (GGT)

Cholinesterase/
Pseudocholinesterase

Angiotensin-Converting
Enzyme
Ceruloplasmin
Ornithine carbamoyl
transferase
G-6-PD
Normal Values
(Enzymes)

Most specific indicator of myocardial damage (AMI)

Not elevated in angina
1. Tanzer-Gilbarg (forward/direct) = pH 9.0, 340nm
2. Oliver-Rosalki/ Rosalki & Hess (reverse/indirect) = most
commonly used method, faster reaction; pH 6.8, 340nm
Inside RBCs
Interferes w/ CK assay
Inhibited by adenosine monophosphate
Activate CK
Do not contain CK
Partially restore lost activity of CK
Reference method for CK
CKI (%) = CK-MB/Total CK x 100
Isoenzymes:
Aldolase A = Skeletal muscles
Aldolase B = WBC, liver, kidney
Aldolase C = brain tissue
Marker for hepatobiliary diseases and infiltrative lesions of the liver
Methods:
1. Dixon and Purdon
2. Campbell, Belfield and Goldberg
Located in the canaliculi of the hepatic cells
Differentates the source of an elevated ALP level
Sensitive indicator of occult alcoholism
Increased:
Obstructive jaundice
Alcoholic hepatitis (most sensitive)
Substrate: gamma-glutamyl-p-nitroanilide
1. Szass
2. Rosalki and Tarrow
3. Orlowski
Monitor effects of relaxants (succinylcholine) after surgery
Marker for organophosphate poisoning (Low CHS)
Methods:
1. Ellman technic
2. Potentiometric
A.k.a. peptidyldipeptidase A or Kininase II
Converts angiotensin I angiotensin II (lungs)
Indicator of neuronal dysfunction (Alzheimers disease CSF)
Ferrooxidase enzyme
For hepatobiliary diseases
Drug induced hemolytic anemia (primaquine, antimalarial drug)
ALP = 30-90 U/L
ACP:
Total ACP (male) = 2.5-11.7 U/L
Prostatic ACP = 0-3.5 ng/mL
AST = 5-37 U/L
ALT = 6-37 U/L
lec.mt 04 |Page | 5

AMS = 60-180 SU/dL (95-290 U/L)

LPS = 0-1.0 U/mL
LDH:
Forward = 100-225 U/L
Reverse = 80-280 U/L
Acute Myocardial Infarction Markers
Myoglobin
Troponin T
Troponin I
CK-MB
AST
LD
Rise
1-3 h
3-4 h
3-6 h
4-8 h
6-8 h
12-24 h
Peak
5-12 h
10-24 h
12-18 h
12-24 h
24 h
48-72 h
Normalize
18-30 h
7 d (10-14
5-10 d
48-72 h
5d
10-14 d
d)
Acute Pancreatitis Markers
Amylase
Lipase
Rise
2-12 h
6h
Peak
24 h
24 h
Normalize
3-5 d
7d
Electrolytes
Electroneutrality
Equal no. of cations and anions
Balance of charges
40-75%
Average water content of the human body
ECF
1/3 of total body water
ICF
2/3 of total body water
Normal plasma
93% water (Plasma: 13% > Whole blood)
7% solutes: (Increased in dehydration)
-Proteins
-Glucose
-NPN
-Lipids
-Ions
Vasopressin deficiency
Excretion of 10-20L H2O everyday
Volume and Osmotic
Sodium
regulation
Potassium
Chloride
Electrolytes
EC = Na+ > Cl- > HCO3- > Ca2+(5th) > iPO4
IC = K+ > Mg2+(4th)
Myocardial rhythm and Potassium
contractility
Calcium
Neuromuscular
Magnesium
excitability
Cofactors (enzyme)
Calcium
Magnesium (CK)
Zinc
Chloride (AMS)
Potassium
ATPase ion pump
Magnesium
Production and use of
Magnesium
ATP from glucose
Phosphate
Acid-base balance
Bicarbonate
Replication of DNA and Magnesium
translation of mRNA
Sodium
Major contributor of osmolality (92%, together w/ Chloride and
Bicarbonate)
lec.mt 04 |Page | 6

Aldosterone
Atrial natriuretic factor
Hypernatremia

Hyponatremia

Thirst

Pseudohyponatremia
(artifactual)
Methods (Na+)

Potassium
Specimen
Considerations (K+)

Hyperkalemia

Hypokalemia

pH and K+
Methods (K+)

Chloride

100 mg/dL glucose = 1.6 mmol/L sodium

Sodium
Potassium = Magnesium
Sodium
Excess water loss
Decreased water intake
Hyperaldosteronism (Conns disease)
Hypothalamic disease (Chronic hypernatremia)
Renal failure
SIADH (increased water retention)
Marked hemolysis (dilutional effect)
<125 mmol/L = severe neuropsychiatric symptoms
Major defense against hyperosmolality and hypernatremia
1-2% water deficit = severe thirst
150-160 mEq/L Na+ = Moderate deficit of water
>165 mEq/L Na+ = Severe water deficit
Hyperlipidemia (turbidity)
Hyperproteinemia
1. FEP
2. AAS
3. ISE = Glass aluminum silicate
4. Colorimetry = Albanese Lein
Concentration in RBC is 105 mmol/L
Reciprocal relationship with H+
0.5% hemolysis = 0.5 mmol/L
Gross hemolysis = 30%
Serum K+ > Plasma K+ by 0.1-0.7 mmol/L because of platelets
(clot)
10-20% in muscle activity
0.3-1.2 mmol/L = mild to moderate exercise
2-3 mmol/L = vigorous exercise; fist clenching
Decreased resting membrane potential incr. contractility lack
of muscle excitability
Decreased renal excretion (Dehydration, renal failure, Addisons
disease)
Acidosis (DM)
Muscle injury
Spironolactone
Increased resting membrane potential arrhythmia
Leads to hypomagnesemia
Vomiting
Diuretics
Cushings syndrome
Alkalosis
Insulin overdose
pH by 0.1 = K+ by 0.2-1.7 mmol/L
Lithium heparin plasma = preferred
1. FEP
2. AAS
3. ISE = Valinomycin gel
4. Colorimetry = Lockhead and Purcell
Chief counter ion of sodium in ECF
lec.mt 04 |Page | 7

Specimen
Considerations (Cl-)
Methods (Cl-)

Hyperchloremia

Hypochloremia

Calcium

3 Forms of Calcium
Vitamin D3
PTH
Calcitonin
Practical considerations
(Ca2+)
Hypercalcemia

Hypocalcemia
Primary hypocalcemia
Secondary
hypocalcemia
Methods (Ca2+)

Chloride methods measure bromide and iodide

Cl- = HCO31. Schales and Schales:
-Mercurimetric titration
-Diphenylcarbazone
-Excess Hg++
-(+) Blue violet
2. Whiterhorn Titration method
-Mercuric thiocyanate
-Reddish complex
3. Ferric perchlorate
4. Cotlove chloridometer
-Coulometric amperometric titration
-Excess Ag++
5. ISE
-Ion exchange membrane
-Tri-n-octylpropylammonium chloride decanol
Renal tubular acidosis
Metabolic acidosis
Diabetes insipidus (Dehydration)
Prolonged diarrhea
Prolonged vomiting (HCl)
Aldosterone deficiency (Na+ = Cl- = K+)
Metabolic alkalosis (HCO3- = Cl-)
Marked hemolysis (dilutional effect)
99% Bones
1% ECF
Absorbed in the duodenum
Absorption is favored at an acidic pH
50% = Free/Ionized/Unbound/Active Calcium
40% = Protein-bound (Albumin)
10% = Complexed with anions
Ca2+ = absorption (intestine) and reabsorption (kidney)
Ca2+ = resorption (bone) and reabsorption (kidney)
Ca2+ = urinary excretion (major net loss of calcium)
Serum = specimen of choice
Albumin (1g/dL) = Ca2+ (0.8 mg/dL)
Acidosis (Ca2+: from Bones Blood)
Cancer
Hyperthyroidism
Milk-alkali syndrome
Tetany
Alkalosis (Ca2+: from Blood Bones)
Acute pancreatitis (Ca2+: binds to damage pancreatic tissues)
Low PTH
Parathyroid gland disease
High PTH
Renal failure ( excretion)
1. Clark Collip precipitation method
-(+) Oxalic acid
-Renal calculi
2. Ferro Ham Chloranilic acid precipitation method
lec.mt 04 |Page | 8

Inorganic Phosphorus

3 Forms of Inorganic
Phosphorus
PTH
Calcitonin
Growth hormone
Practical considerations
Hyperphosphatemia
Hypophosphatemia
Methods (iPO4)

Magnesium

3 Forms of Magnesium
PTH
Aldosterone (&
Thyroxine)
Hypermagnesemia
Hypomagnesemia
Methods (Mg2+)

-(+)Chloranilic acid
3. Colorimetric = Ortho-Cresolphthalein complexone dyes
-Dye: Arzeno III
-8-hydroxyquinoline = chelates (inhibits) Mg2+
4. EDTA titration method (Bachra, Dawer and Sobel)
5. AAS = Reference method
6. ISE = Liquid membrane
7. FEP
85% Bones
15% ECF (iPO4)
Maximally absorbed in the jejunum (Ca2+: duodenum)
Trancellular shift: Once absorbed inside cells, it no longer comes
out used for energy production
Dirunal variation: late morning, evening
Organic phosphate = principal anion within cells
Inorganic phosphate = part of the blood buffer (Measured in the
clin.lab.)
55% = Free
35% = Complexed with ions
10% = Protein-bound
PO4 = Ca2+
PO4 = Ca2+
PO4 (renal reabsorption)
Fasting is required (Nonfasting: PO4)
Hypoparathyroidism
Renal failure
Hypervitaminosis D
Alcohol abuse = most common cause
Primary hyperparathyroidism
Avitaminosis D (Rickets, Osteomalacia)
Most accurate: unreduced phosphomolybdate formation (340nm)
1. Fiske Subbarow Method (Ammonium molybdate method)
-Reducing agents: Pictol, Elon, Senidine, Ascorbic acid
-(+) Phosphomolybdenum blue
53% Bones
46% Muscles and soft tissues
1% Serum and RBC
Vasodilator
55% = Free/Ionized/Physiologically active
30% = Protein-bound
10% = Complexed with ions
Mg2+ = Ca2+ = PO4
Mg2+ = K+ = Na+
Addisons disease
Chronic renal failure
Acute renal failure
Chronic alcoholism
1. Calmagite
-(+) Reddish-violet complex
2. Formazen dye method
-(+) Colored complex
3. Magnesium Thymol blue method
lec.mt 04 |Page | 9

Bicarbonate
Chloride shift
Anion Gap
Increased AG

Decreased AG

Cystic Fibrosis
(Mucoviscidosis)

Pilocarpine
Gibson & Cooke
pilocarpine
iontophoresis
Iron

Methods (Iron)

Increased iron
Decreased iron
TIBC
UIBC
% Transferrin
Saturation

-(+) Colored complex

4. AAS = reference method
5. Dye-lake Method
-Titan Yellow dye (Clayton Yellow or Thiazole yellow)
90% of the total CO2
HCO3- diffuses out of the cell in exchange for Cl - to maintain ionic
charge neutrality w/in the cell
Difference between unmeasured anions and unmeasured cations
QC for ISE
Uremia/renal failure
Ketoacidosis
Lactic acidosis
Methanol poisoning
Ethanol poisoning
Ethylene glycol poisoning
Salicylate poisoning
Hypoalbuminemia
Hypercalcemia
Hyperlipidemia
Multiple myeloma
Defective gene: Cystic fibrosis transmembranous conductance
regulator (Chromosome 7)
Miconeum ileus (Infants)
Foul-smelling stool
URT infection
Na+ and ClSweat inducer
Reference method (Sweat sodium and chloride)
Prooxidant
3-5g = Total body iron
Ferrous = Hgb
Ferric = Transferrin and Ferritin
1. Colorimetric = HCl and Ferrozine
-(+) Blue color
2. Anodic stripping voltammetry
Hemochromatosis
Viral hepatitis
Non-IDA
IDA
Malnutrition
Chronic infection
UIBC + Serum Iron
Increased: IDA, hepatitis, iron-supplemented pregnancy
Decreased: Non-IDA, nephrosis
TIBC Serum iron
Measure of reserve iron binding capacity of transferrin
Index of iron storage
Increased: Iron overdose, hemochromatosis, sideroblastic anemia
Decreased: IDA (lowest), malignancy, chronic infection
lec.mt 04 |Page | 10

Transferrin
Note

Normal Values
(Electrolytes)

Regulation of Acid-Base
balance
20:1
4:1
Expanded HendersonHasselbalch equation
Chloride-isohydric shift
pCO2
pO2
Metabolic Acidosis

TIBC (g/dL) x 0.70 = mg/dL

Sodium 1/ Potassium
Potassium 1/ Hydrogen ion
Potassium Magnesium
Magnesium Calcium
Calcium 1/ Inorganic phosphate
Chloride 1/ Bicarbonate
Sodium:
Serum = 135-145 mmol/L
[Critical: 160 mmol/L and 120 mmol/L]
CSF = 136-150 mmol/L
Potassium:
Serum = 3.5-5.2 mmol/L
[Critical: 6.5 mmol/L and 2.5 mmol/L]
Chloride:
Serum = 98-107 mmol/L
Sweat = 5-40 mmol/L [Critical: >65 mmol/L]
Calcium:
Total = 8.6-10 mg/dL (adult) and 8.8-10.8 mg/dL (child)
Ionized = 4.6-5.3 mg/dL (adult) and 4.8-5.5 mg/dL (child)
[Critical: <7.5 mg/dL]
Inorganic Phosphate:
Adult = 2.7-4.5 mg/dL
Child = 4.5-5.5 mg/dL
Magnesium:
Serum = 1.2-2.1 mEq/L
Anion Gap:
w/ K+ = 10-20 mmol/L
w/o K+ = 7-16 mmol/L
Iron:
Male = 50-160 g/dL
Female = 45-150 g/dL
TIBC:
Adult = 245-425 g/dL
>40 y.o. = 10-250 g/dL
NB and Child = 100-200 g/dL
% Transferrin Saturation = 20-50%
Blood Gases and pH
Lungs and Kidneys
CO2 + H2O <--(Carbonic anhydrase)--> H2CO3
H2CO3 <-------(Carbonic anhydrase)--> H+ + HCO3HCO3-: H2CO3 ratio
HPO4: H2PO4 ratio
pH = 6.1 + log [Total CO2 (pCO2 x 0.03)]
pCO2 x 0.03
Buffering effect of hemoglobin
Index of efficiency of gas exchange
Increased: Barbiturates, morphine, alcohol, heparin (12-15%)
Reflects the availability of the gas in blood but not its content
Excessive O2 supply acidosis
Causes:
lec.mt 04 |Page | 11

Metabolic Alkalosis

Respiratory Acidosis

Respiratory Alkalosis

Full compensation
Partial compensation
Buffer base
Methods for Blood
Gases and pH
Factors affecting Blood
gases & pH
measurements

Methods
(Blood gases & pH)

Whole blood total CO2

-Bicarbonate deficiency
-DKA (normochloremic acidosis)
-Renal failure
-Diarrhea (HCO3-)
Compensation: Hyperventilation
Compensated: HCO3- + pCO2 + pH <7.4
Causes:
-Bicarbonate excess
-Vomiting (Cl-)
-Hypochloremia
-Hypokalemia
Compensation: Hypoventilation
Compensated: HCO3- + pCO2 + pH >7.4
Causes:
-CO2 excess (Hypoventilation)
-COPD
-Drug overdose (morphine, barbiturates, opiates)
Compensation: Bicarbonate retention
Compensated: HCO3- + pCO2 + pH <7.4
Causes
-CO2 loss (Hyperventilation)
Compensation: Bicarbonate excretion
Compensated: HCO3- + pCO2 + pH >7.4
pH normal range
pH near normal
All forms of base that will titrate hydrogen ions
Specimen: Arterial blood
Blood gas analyzers: meas. pH, pCO2, pO2
For every 1OC above 37OC:
pH by 0.015
pO2 by 7%
pCO2 by 3%
Bacterial contamination: consume O2 (pO2)
Excess heparin (acid MPS) = pH
Air exposure (bubbles):
pO2 = 4 mmHg/2mins
pCO2 = 4 mmHg/2mins
1. Gasometer
a. Van Slyke
b. Natelson
-Mercury: produce vacuum
-Caprylic alcohol: anti-foam reagent
-Lactic acid
-NaOH
-NaHSO3
2. Electrodes
a. pH = potentiometry
-Silver-silver chloride electrode (Reference electrode)
-Calomel electrode [Hg2Cl2] (Reference electrode)
b. pCO2 = Severinghaus electrode (potentiometry)
c. pO2 = Clark electrode (polarography-amperometry)
Dissolved CO2 + H2CO3 + HCO3lec.mt 04 |Page | 12

Transcutaneous
electrodes
Blood gas QC
Normal Values
(Blood gases and pH)

Continuous monitoring of pO2

Directly placed on the skin
Min. requirement:
-1 sample every 8 hours
-3 levels of control (acidosis, normal, alkalosis) every 24 hours
pH = 7.35-7.45
pCO2 = 35-45 mmHg
Total CO2:
WB arterial = 19-24 mmol/L
WB venous = 22-26 mmol/L
HCO3- = 21-28 mEq/L
pO2 = 81-100 mmHg
[Hypoxemia:]
-Mild (61-80 mmHg)
-Moderate (41-60 mmHg)
-Severe (40 mmHg or less)
O2 saturation = 94-100%

lec.mt 04 |Page | 13