Professional Documents
Culture Documents
Enzymatic
1. Glucose Dehydrogenase- glucose is converted by glucose
dehydrogenase into gluconolactone with the simultaneous
conversion of NAD into NADH; a chromophore is produced at the
end of the series of reactions
2. Glucose Oxidase- this is the most specific enzyme for D
Glucose; involves the conversion of glucose into gluconic acid
with H2O2 production
Colorimetric Glucose Oxidase- involves a second reaction using
Peroxidase
Saifer Gernstenfield- uses O- dianisidine as chromogen
Trinder- used phenylphenazone as chromogen
LABORATORY DIAGNOSIS
1. RBG- uses randomly collected samples; especially useful in cases of
hypoglycemia, insulin shock, and hyperglycemic coma; a value 200
mg/dl with signs and symptoms can be used to diagnose diabetes
2. FBG- requires an 8- hour fast; useful for diabetes diagnosis
Analyte
Fasting Blood
Glucose
Random Blood
Glucose
OGCT
OGTT (Fasting)
OGTT (1st hour)
OGTT (2nd hour)
OGTT (3rd hour)
HbA1c
REFERENCE INTERVALS
Conventional Unit Conversion Factor
70-100 mg/dl
0.056
SI Unit
3.9-5.6 mmol/l
0.056
<130 mg/dl
DM 70-100 mg/dl
GDM <95 mg/dl
DM <140 mg/dl
GDM <180 mg/dl
DM <140 mg/dl
GDM <155 mg/dl
GDM <140 mg/dl
4-5.6%
0.056
0.056
0.056
0.056
0.056
0.01
<7.2 mmol/l
3.9-5.6 mmol/l
<5.3 mmol/l
<7.8 mmol/l
<10.0 mmol/l
<7.8 mmol/l
<8.6 mmol/l
<7.8 mmol/l
0.04-0.056
AMMONIA
- byproduct of protein catabolism
- converted to urea
- increased in liver disease, Reyes Syndrome
- Analytical Techniques
Isothermal Diffusion
Enzymatic- uses the enzymatic reaction of ammonia with
ketoglutarate in the presence of glutamine dehydrogenase
AMINO ACIDS
- basic units of proteins
- 20 biologically significant
- essential vs nonessential
- amphoteric
- aminoacidurias may be overflow or due to inborn errors of metabolism
- Amino Acidopathies
Phenylketonuria (PKU)- inherited deficiency of phenylalanine
hydroxylase needed for the conversion of phenylalanine to tyrosine;
increased phenylalanine and phenylpyruvic acid in plasma and urine; diet
should lack phenylalanine to prevent cerebral damage
MSUD- deficiency of branched chain alpha keto acid dehydrogenase
that results to the accumulation of the branched chain amino acids
isoleucine, leucine, and valine
Transient Neonatal Tyrosinemia- most common amino acid disorder in
the neonatal period; due to an imbalance of elevated serum tyrosine
levels from protein intake and delayed maturation of the hepatic enzyme
parahydroxyphenylpyruvate dioxygenase
Secondary Aminoaciduria- may occur in cirrhosis and chronic liver
diseases
Renal Aminoaciduria- due to diminished tubular reabsorption; may be
acquired due to kidney damage or an inborn error of metabolism
Ammonia
REFERENCE INTERVALS
Conventional Unit Conversion Factor
8-23 mg/dl
0.357
F- 0.5-1.0 mg/dl
M- 0.6-1.2 mg/dl
F- 0.2-0.7 mg/dl
M- 0.1-0.4 mg/dl
F- 2.7-7.3 mg/dl
M- 4.0-8.5 mg/dl
<114 ug/dl
88.4
76.25
0.059
0.59
SI Unit
2.9-8.2 mmol/l
F- 44-88 umol/L
M- 53-106 umol/L
F- 15-53 umol/l
M- 8-31 umol/l
F 0.16-0.43
mmol/L
M 0.24-0.51
mmol/L
< 67 umol/L
PROTEINS
Definition- polymers of amino acids produced by living cells
Composed of amino acids in varying numbers and sequences
Approximately 16% nitrogen content
Functions:
1. maintenance of plasma colloid osmotic pressure
2. transport of water- insoluble substances
3. antibodies
4. hormone structure
5. nutrients for building materials
6. catalyst
7. clotting
Classifications
A. Structure- primary, secondary, tertiary, quarternary
B. Shape- globular, fibrous
C. Solubility
D. Composition- simple, conjugated
E. Electrophoretic Mobility
SPECIFIC PLASMA PROTEINS
9. Nephelometry
10.
Immunoassay
Analyte
Total Protein
Albumin
Globulin
A/G Ratio
Serum Protein
Electrophoresis
REFERENCE INTERVALS
Conventional Unit Conversion Factor
6.0-7.8 g/dl
10
3.2-4.5 g/dl
10
2.3-3.5 g/dl
10
1.3-3.0
% Total Protein
Albumin- 52-65%
0.01
1- 2.5-5%
0.01
2- 7-13%
0.01
- 8-14%
0.01
- 12-22%
0.01
SI Unit
60-78 g/l
32-45 g/l
23-35 g/l
1.3-3.0
Fraction Total
Protein
0.52-0.65
0.025-0.055
0.07-0.13
0.08-0.14
0.12-0.22
CLASSIFICATION
1. Fatty acids- long chain hydrocarbons
- Short chain, medium chain or long chain
- Saturated, monounsaturated or polyunsaturated
- Even or odd numbers of carbons
- Cis or Trans configuration
2. Glycerol esters
TAG- prevalent glycerol ester in the plasma and adipose tissues; has 3
fatty acids attached to glycerol
Phospholipids- has a phosphoric acid head group in one of the carbons
of glycerol and 2 fatty acids
Over 90% phosphatidylcholine and sphingomyelin, 3-6%
ethanolamine and serine, and 4-9% lysophosphatidylcholine
3. Sterol Derivatives
Cholesterol- source of primary and secondary bile acids, steroid
hormones and Vitamin D; Has CPPP nucleus; 60-70% esterified and
30-40% unesterified
4. Sphingosine Derivatives (Sphingolipids)
Ceramide- with fatty acid bound to sphingosine; CNS cell membrane
structure; sphingomyelin, galactosylceramide, glucosylceramide
Terpenes- with 5 branched chain units; intermediates in the production
of cholesterol; includes the fat- soluble vitamins
MAJOR LIPID CONSTITUENTS IN THE PLASMA
1. Cholesterol
2. Phospholipids
3. TAG
LIPOPROTEINS
Lipoproteins transport all the essential cholesterol and lipids in the plasma;
for the packaging, solubility, and metabolism of lipids
Apolipoproteins- the protein moiety of lipoproteins; influences enzymes in
lipid metabolism; helps bind lipoproteins to cell surface receptors
General Lipoprotein Structure
APOLIPOPROTEINS
1. Apo A- Apo A1 and Apo A2
2. Apo B- Apo B48 and Apo B 100
REFERENCE INTERVALS
Conventional Unit Conversion Factor
400-800 mg/dl
0.01
150-250 mg/dl
0.026
10-190 mg/dl
0.0113
150-380 mg/dl
0.01
9-15 mmol/l
1
SI Unit
4-8 g/l
3.88-6.47 mmol/l
0.11-2.15 mmol/l
1.5-3.8 g/l
9-15 mmol/l
ENZYME MEASUREMENTS
Concentration- can be determined by immunologic techniques
Activity- evidenced by increase in product formation, decrease in amount of
substrate, or changes in coenzyme concentration
Methods
1. Fixed Time/ End Point
2. Kinetic/ Continuous Monitoring
Units
1. International Units
2. Katal- SI Unit
Enzymes of Clinical Significance and their EC Nomenclature
1. Lactate Dehydrogenase (LD)
EC 1.1.1.27
2. Alanine Aminotransferase (ALT)
EC 2.6.1.2
3. Aspartate Aminotransferase ( AST) EC 2.6.1.1
4. Creatine Kinase (CK)
EC 2.7.3.2
5. Alkaline Phosphatase (ALP)
EC 3.1.3.1
6. Acid Phosphatase (ACP)
EC 3.1.3.2
7. Amylase (AMS)
EC 3.2.1.1
8. Lipase (LPS)
EC 3.1.1.3
9. Gamma Glutmayl Transferase (GGT)
EC 2.3.2.2
10.
5 Nucleotidase (5 N)
EC 3.1.3.5
11.
Leucine Aminopeptidase (LAP)
EC 3.4.11.1
12.
Acetylcholinesterase (ACE)
EC 3.4.15.1
13.
Pseudocholinesterase (PCHE)
EC 3.1.1.8
14.
Aldolase (ALD)
EC 4.1.2.13
Creatine Kinase
Abbreviation: CK, CPK
Systematic Name: ATP: Creatine N-phosphotransferase
Clinical Significance: MI, skeletal muscle disorders
Functions: ATP regeneration
Reaction Catalyzed
Lactate Dehydrogenase
Abbreviation: LD, LDH
Systematic Name: L- Lactate: NAD+ Oxidoreductase
Clinical Significance: MI, Liver Disorder, Hemolysis, Megaloblastic
Anemia, CA
Functions: Hydrogen Transfer
Reaction Catalyzed
Aspartate Aminotransferase
Abbreviation: AST, SGOT
Systematic Name: L- Aspartate: 2- Oxaloglutarate Aminotransferase
Assay Methods
1. Karmen Method
Alanine Aminotransferase
Abbreviation: ALT, SGPT
Systematic Name: L- Alanine: 2- Oxaloglutarate Aminotransferase
Clinical Significance: Liver Disorder,
Functions: Synthesis & Degradation of Amino Acids in Intermediary
Metabolism
Reaction Catalyzed
Assay Methods
Alkaline Phosphatase
Abbreviation: ALP
Systematic Name: Orthophosphoric monoester phosphohydrolase
(alkaline)
Clinical Significance: Bone Disorder, Liver Disorder
Functions: Liberates inorganic phosphate from an organic phosphate
ester
Reaction Catalyzed
Assay Methods
1. Bowers and McComb
Acid Phosphatase
Abbreviation: ACP
Systematic Name: Orthophosphoric monoester phosphohydrolase
(acid)
Clinical Significance: Prostate CA
Functions: Liberates inorganic phosphate from an organic phosphate
ester
Reaction Catalyzed
Glutamyltransferase
Abbreviation: GGT, GGTP
Systematic Name: (5-Glutamyl)peptide: amino acid-5glutamyltransferase
Clinical Significance: Liver Disorder
Functions: Peptide and protein synthesis, regulation of tissue
glutathione levels, and transport of amino acids across cell membranes
Reaction Catalyzed
Assay Methods
Amylase
Abbreviation: AMY, AMS
Systematic Name: 1,4 D-Glucan Glucanohydrolase
Clinical Significance: Acute Pancreatitis
Functions: Catalyzes the breakdown of starch and glycogen
Reaction Catalyzed
Isoenzymes: P and S
Assay Methods
1. Amyloclastic
2. Saccharogenic
3. Chromogenic
4. Continuous Monitoring
Lipase
Abbreviation: LPS
Systematic Name: Triacylglycerol Acylhydrolase
Clinical Significance: Acute Pancreatitis
Functions: Catalyzes the partial hydrolysis of dietary TAG
Reaction Catalyzed
Assay Methods
1. Titrimetric
2. Turbidimetric
Glucose-6-Phosphate Dehydrogenase
Abbreviation: G6PD, G6PDH
Systematic Name: D- Glucose-6-Phosphate: NADP+ 1- Oxidoreductase
Clinical Significance: Drug- induced Hemolytic Anemia
Functions: NADPH production in the Pentose Phosphate Shunt
Assay Methods
Analyte
Amylase
Creatine Kinase
Glucose 6
Phosphate
Dehydrogenase
Glutamyltransfer
ase
Lactate
dehydrogenase
Leucine
aminopeptidase
REFERENCE INTERVALS
Conventional Unit Conversion Factor
SI Unit
16-120 Somogyi
1.85
30-220 U/L
units/dl
M- 55-170 U/L
1
M- 55-170 U/L
F- 30-135 U/L
F- 30-135 U/L
12002000
1
12002000 U/L
mU/mL packed
packed
erythrocytes
erythrocytes
540 U/L
1
540 U/L
(lactate
pyruvate) 100
190 U/L
Male 80200
U/mL (Goldbarg
Rutenberg)
Female 75185
100190 U/L at
37 C
0.24
19.248 U/L
1844.4 U/L
Lipase
5 Nucleotidase
Acid phosphatase
Alkaline
phosphatase
Aspartate amino
transferase
Alanine amino
transferase
U/mL (Goldbarg
Rutenberg)
14280 mU/mL
01.6 units at 37
C
0.130.63 U/L at
37 C
20130 U/L at 37
C
833 U/L at 37 C
436 U/L at 37 C
1
1
14280 U/L
01.6 units at 37
C
2.210.5 U/L at
37 C
20130 U/L at 37
C
833 U/L at 37 C
436 U/L at 37 C
16.67
1
1.
2.
3.
1.
2.
Enzymes
Angiotensin Sensitivity Test- first marker developed; obsolete
Lactate Dehydrogenase- not specific but isoenzymes LD1 and
LD2 are used; 2-4 times elevation from the upper limit of normal
that peaks at 48- 72 hours from chest pain onset and goes back
to baseline within 10 days; flipped LD pattern
Creatine Kinase- most specially CKMB
CK MB- rises 4-6 hours from the onset of chest pain, peaks within
12- 24 hours, and returns to baseline in 48- 72 hours
CK MB Mass/ CK Activity of >3 is associated with AMI.
Cardiac Proteins
Myoglobin- released upon cardiac damage; small and rapidly
cleared by the kidneys so it cannot be used as a long term
marker; not specific for AMI; rises within 1-4 hours from the onset
of chest pain, peaks within 6-9 hours, but declines to baseline
within 18- 24 hours; it cannot be used for those with renal
disease because of slower clearance; can be used to detect reinfarction if it elevates again in the next few days
Troponin Complex- includes Troponin C (least cardiac-specific),
Troponin I and Troponin T; not detectable in the serum of healthy
individuals
Troponin T- allows for both early and late AMI diagnosis;
increases a few hours after the onset of chest pain and peaks by
2.
3.
4.
5.
1.
2.
3.
4.
5.
1.
Analyte
Creatine Kinase
CK MB
Myoglobin
Troponin I
Lactate
dehydrogenase
Aspartate amino
transferase
REFERENCE INTERVALS
Conventional Unit Conversion Factor
M- 55-170 U/L
1
F- 30-135 U/L
<6% of total
<90 g/L
1
(lactate
pyruvate) 100
190 U/L
833 U/L at 37 C
SI Unit
M- 55-170 U/L
F- 30-135 U/L
<6% of total
<90 g/L
00.1 g/L
100190 U/L at
37 C
833 U/L at 37 C
Liver Disorders
1. Jaundice- yellowish discoloration of the skin and sclera due to
increased bilirubin; prehepatic, hepatic, or posthepatic
2. Cirrhosis
3. Hepatitis
4. Biliary Obstruction
5. Tumors
6. Reyes Syndrome
7. Alcohol and Drug Related
Assessment of Liver Function
1. Bilirubin
2.
3.
4.
5.
6.
7.
Ehrlichs
Van Den Bergh Reactions
Evelyn Malloy Method
Jendrassik Grof
Icterus Index
Direct Spectrophotometry
Urobilinogen
Bile Acids
Enzymes- AST, ALT, ALP, 5N, LAP, LDH, GGT
Test Measuring Synthetic Activity- PT, APTT, Albumin, Globulin,
A/G
Ammonia
Older Tests: Vitamin K Response Test, Hippuric Acid Synthesis
Test
Analyte
Ammonia
Bile acids
Bilirubin
Direct
(conjugated)
Indirect
(unconjugated)
Total
Newborns total
Glutamyltransfer
ase
Lactate
dehydrogenase
Leucine
aminopeptidase
REFERENCE INTERVALS
Conventional Unit Conversion Factor
SI Unit
20120 g/dL
0.5872
1270 mol/L
(diffusion)
4080 g/dL
2347 mol/L
(enzymatic
method)
728 mol/L
1248 g/dL
(resin method)
0.33 mg/dL
10
330 mg/L
<0.3 mg/dL
0.11 mg/dL
0.11.2 mg/dL
112 mg/dL
17.10
<5 mol/L
217 mol/L
221 mol/L
17205 mol/L
540 U/L
540 U/L
(lactate
pyruvate) 100
190 U/L
Male 80200
U/mL (Goldbarg
Rutenberg)
Female 75185
100190 U/L at
37 C
0.24
19.248 U/L
1844.4 U/L
Isocitric
dehydrogenase
5 Nucleotidase
Alkaline
phosphatase
Aspartate amino
transferase
Alanine amino
transferase
U/mL (Goldbarg
Rutenberg)
50240 U/mL at
25 C (WolfsonWilliamsAshman
units)
01.6 units at 37
C
20130 U/L at 37
C
833 U/L at 37 C
436 U/L at 37 C
0.0166
0.834.18 U/L at
25 C
01.6 units at 37
C
20130 U/L at 37
C
833 U/L at 37 C
436 U/L at 37 C
Artifactual Hyperkalemia
Seen in hight platelet count, prolonged tourniquet application,
hemolysis, delayed separation, refrigeration
! Very high potassium levels can stop the heartbeat.
Analytical Techniques
! Serum vs Plasma
1. ISE
2. FES
3. Spectrophotometric: Lockhead and Purcell, Turbidimetric
4. AAS
CHLORIDE
Most abundant ECF anion
Also regulates osmotic pressure and water balance together with
sodium
Hypochloridemia
Metabolic alkalosis, respiratory acidosis
Hyperchloridemia
Metabolic acidosis, respiratory alkalosis
Analytical Techniques
1. Mercurimetric Titration- Schales and Schales
2. Mercuric Thiocyanate Methods- Skeggs Modification
3. Coulometric- Amperometric Titration
4. ISE
! Sweat Chloride Determination
CALCIUM
Hypercalcemia
Hyperparathyroidism, multiple myeloma, TB, antacids
Analytical Techniques
Total Calcium
1. Colorimetric- Clark and Collip
2. EDTA Titration
3. Spectrophotometric- O- Cresolphthalein, Alizarin, Arsenazo III
4. AAS
Ionized
1. Colorimetric
2. ISE
PHOSPHORUS
Found in bones, muscles, and ECF
Also influenced by PTH, Calcitonin, and Vit D
For structural support, energy generation, and storage
Analytical Techniques
1. Spectrophotometric- Fiske and Subarrow
MAGNESIUM
2nd most abundant cation in the ICF
4th most abundant cation in the body
As an activator of various enzymes
Forms in the Blood: Free or Ionized (2/3) and Protein Bound (1/3)
Analytical Techniques
1. AAS
2. EFP
3. Spectrophotometric- Calmagite Green, Formazan Dye, Methylene Blue
and others
4. Colorimetric- Titan Yellow
BICARBONATE (Total CO2)
2nd most abundant ECF anion
Chloride Shift
Acid- base balance
Analytical Techniques
1. Acid Titration and pCO2 Electrode
2. Akalinization and En
Analyte
Chloride
Calcium
Ionized
Total
Bicarbonate
Magnesium
Osmolality
Phosphorus,
inorganic
Potassium
Sodium
REFERENCE INTERVALS
Conventional Unit Conversion Factor
SI Unit
95103 mEq/L
1
95103 mEq/L
24 hr urine 140
1
140250
250 mEq/day
mmol/day
Sweat 460
1
mEq/L
460 mmol/L
44.8 mg/dL
0.2500
11.2 mmol/L
22.4 mEq/L
0.5000
3058% of total
0.01
0.300.58 of total
Plasma 2128
1
Plasma 2128
mmol/L
mmol/L
1.32.1 mEq/L
0.5000
0.651.05 mmol/L
1.83 mg/dL
0.4114
0.741.23 mmol/L
24 hr urine 68.5
0.5000
34.3 mmol/day
mEq/day
280295
1
280295
mOsm/kg
mOsm/kg
Adults 2.34.7
0.3229
0.741.52 mmol/L
mg/dL
1.292.26 mmol/
Children 47
mg/dL
3.85 mEq/L
1
3.85 mmol/L
24 hr urine 4080
1
4080 mmol/day
mEq/day
136142 mEq/L
1
136142 mmol/L
24 hr urine 75
1
75200 mmol/day
200 mEq/day
Sweat 1080
1
1080 mmol/L
mEq/L
pCO2
pO2
Bicarbonate
Oxygen
Saturation
(arterial)
7.367.41
(venous)
3540 mm Hg
(arterial)
4045 mm Hg
(venous)
95100 mm Hg
(arterial)
21-28 mEq/L
94100%
7.367.41
0.1333
4.75.3 kPa
5.36 kPa
0.1333
12.713.3 kPa
1
0.01
21-28 mmol/L
Fraction
saturated:
0.941
TOXICOLOGY
Basic Techniques
Immunochemical Methods- homogeneous immunoassays; EMIT and FPIA
Chromatographic Techniques- TLC, HPLC, GC- MS
DRUGS OF ABUSE
Cocaine
derived from coca plant
prevalent form of cocaine is called crack a free-base form that passes rapidly
across the nasal membranes
half-life is 1-2 hours where as parent compound and its metabolites cleared from
the body within 2 days
normal administration of cocaine is nasal (inhalation or snorting)
Opiates (Morphine, Codeine, Heroin)
Clinical Significance
Morphine powerful analgesic, binding to receptors in the CNS; used in
treating acute CHF
Codeine - mild analgesic and as an antitussive
Heroin
- induces a pleasant, euphoric state and is highly addictive both
physically and physiologically
Amphetamines
It causes euphoria and increased mental alertness.
Competitive inhibitors of the enzyme monoamine oxidase which inactivates
adrenergic neurotransmitters by oxidatively removing their amino groups.
3,4-methylenedioximethamphetamine (MDMA or ecstacy) a derivative of
methamphetamine
CNS and respiratory stimulation and sympathomimetic activity
Loss of weight as a result of an anorectic effect
Psychic stimulation and excitability, temporary increase in mental and physical
activity
Benzodiapines
Minor tranquilizers
Class III block repolarizing potassium currents, increasing the length of the
action potential
Class IV verapamil, slow calcium influx resulting in action potential
prolongation
DIGOXIN the digitalis glycosides are used to treat atrial arrhythmias atrial
flutter and fibrillation; rapid onset of action
Digoxin: half life 35-40 hrs; 0.5-2 ng/ml
Digitoxin: half life 4-6 days; 9-25 ng/ml
PROCAINAMIDE (Pronestyl) Class I antiarrhythmic drug used in treating
supraventricular or ventricular arrhythmias
Half life app 3.5 hrs; 4-10 ng/ml
QUINIDINE - Class I antiarrythmic; half-life is 5-12 hours; 2.3-5 mcg/ml
LIDOCAINE (Xylocaine) Class I antiarrythmic, can also be used as local
anesthetic; acute control and prevention of ventricular arrhythmias after
myocardial infraction; half-life is 2 hours; 1.2-5.5 mcg/ml
PROPRANOL- Class II anti-arrhythmic beta-receptor blocking drug; antagonizes
the effects of epinephrine on the heart, arteries, and arterioles of skeletal
muscles, and on the bronchus; used to treat sinus tachycardia, atrial
tachycardia and ventricular arrhythmias; toxic effects include bradycardia,
arterial insufficiency, hypotension, AV block, nausea, vomiting, pharyngitis,
bronchospasm, and thrombic thrombocytopenic purpura; half app 3 hrs; 50-100
ng/ml
AMIODARONE- Class III anti-arrhythmic; oral loading dose is 1200-1600mg/day
with a maintenance dose of 200-400mg/day; toxic effects include bradycardia,
heart block, fatal pulmonary fibrosis, hepatitis, visual field disturbances,
photodermatitis, and mainly hypothyroidism but sometimes hyperthyroidism;
half life rapid- 3-10 days, slow- 25-110 days; 1-2.5 mcg/ml
VERAPAMIL- Class IV anti-arrhythmic drug; oral dose is 120-480mg/day in three
to four divided doses; toxic effects include hypotension, ventricular fibrillation,
constipation, and peripheral edema; half life 2-8 hours; 80-400 ng/ml
Anticonvulsants
Used in the treatment of seizure disorders
PHENOBARBITAL- long-acting barbiturate, used in the treatment of generalized
tonic-clonic seizures and simple partial seizures with motor or somatosensory
symptoms for anxiety and insomnia; toxic side effects include nystagmus,
ataxia, stupor, respiratory depression, coma and hypotension; half life 4-6 days;
15-30 ug/ml
PHENYTOIN (DILANTIN)- for generalized tonic-clonic, simple partial, and complex
partial seizures; toxic side effects include nystagmus, ataxia, stupor, and coma;
half life app 24 hrs; 10-20 ug/ml