Clinical Chemistry

Lecture / PPT based

Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
QUALITY CONTROL • Involves the proficiency testing program
• System of ensuuring accuracy and which provides samples of unknown
precision by including quality control concentration of analytes to
reagents in every series of participating laboratories.
measurements. • Important in maintaining long-term
• A testing design to assess the “health” of accuracy of analytical methos.
an analytical method. LIST OF NATIONAL REFERENCE LABORATORIES
• Involvs the process of monitoring
analytical processess and detects errors. National Kidney Hematology
Transplant Institute
PARAMETERS OF QUALITY CONTROL (NRTI)
1. SENSITIVITY – ability of the analyte to Regional Institute of Microbiology &
detect even the smallest concentration Tropical Medicine (RITM) Parasitology
2. SPECIFICITY – detects only the analyte San Lazaro Hospital (SLH) SACCL
of interest Lung Center of the Chemistry
3. ACCURACY – closeness to the true Philippines (LCP)
value. East Avenue Medical Toxicology and other
4. PRECISION OR REPRODUCTABILITY Center (EAMC) Substance of Abuse
5. PRACTICABILITY – easily prepared NRL Worldwide – Australia
6. RELIABILITY – the accuracy & precision
is still maintained VARIATIONS OF ERRORS
7. DIAGNOSTIC SENSITIVITY – detects RANDOM ERROR
who/what is positive; only measures • Used for varying differences in repeated
the positive pxs. tests
8. DIAGNOSTIC SPECIFICITY – detects A. Mislabeling – never pre-label samples
negative results/patients B. Pipetting errors
Formula: C. Improper mixing of sample and reagents
D. Voltage Fluctuations – all machines are
generate and convertedby AVR; always
do re-testing in case of fluctuations
E. TemperatureFlctuations – reagents
must be stored 2-8°C
F. Dirty Optics
KINDS OF QUALITY CONTROL
Intralab Quality Control (QC) SYSTEMATIC ERROR
• Within the laboraory • Problem with the machine and system
• Involves analyses of control samples itself
together with the patient specimens A. Improper Calibration
• Detects both rndom and systematic B. Detoriation of Reagents
errors C. Contaminated Solutions
• Important for the daily monitoring of D. Sample instability/Unstable Reagent
accuracy and precision methods Blanks
E. Changes in standard materials
Interlab Quality Control (QC) F. Leaky ion selective electrode
• Mandaed by different national reference G. Diminishing lamp power
laboratories (NRLs) H. Incorrect instrument setting

Cuzzamu 1
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT
I.Incorrect sample and reagent volume
J.Poorly written procedures
CONSTANT ERROR
• Refers to a difference between the
target value and assay value.
PROPORTONAL/SLOPE/PERCENT ERROR
• Results to greater deviation from the
target value due to higher sample
concentration.
INTERPRETATION OF QUALITY CONTROL
1. Trend
2. Shift
3. Westgard Multi-rules
• Warning Rules
• Reject Rules
4. Outliers
STATISTICS
• The science of gathering, analyzing,
interpreting and presenting data.
A. Mean
B. Median
C. Mode
D. Standard Deviation
E. Coefficient of variation
F. Variance
TYPES OF QUALITY CONTROL CHART
GAUSSIAN CURVE (BELL-SHAPED CURVE)
• Occrs when the data set can be
accurately described by the SD and
Mean
• Data elements are centered around the
mean with most elements close to the
mean
• Shift in mean = ACCURACY PROBLEM
• Increase in SD = PRECISION PROBLEM
CUMULATIVE SUM GRAPH (CUSUM
• Gives the earliest indication of
systematic errors (trend) and used
within the 1-3s rule
• Very sensitive to small, persistent errors
that commonly occur in low modern-
calibration frequency analyzer.
YOUDEN PLOT
• Used to compare results obtained on a
high and low control serum for different
laboratories.
CUZZAMU, NIKKA AIRA B [BSMT-4A]
SHEWART LEVEY-JENNINGS CHART
• Also known as “Dot Chart”
• Most widely sed in clinical laboratories
• Detects both random and systematic
errors
TREND
• Inicates a gradual loss of reliability in the
system (e.g repetition of values)
Causes:
1. Detoriation of reagents
2. Detoriation of the instrment light source
3. Gradual accumulation of debris in the
sample./reagent tubing or on electrode
surfaces
*QC materials not flexible if the results on the
machine showed the same results over and over.
SHIFT
• Represents a sudden change in the test
performance.
Causes:
1. Inaccurate calibration/re-calibration
2. Change in reagent formulation/reagent
lot
3. Major instrument maintenance
4. Failure in sampling system/reagent
dispense system
OUTLIER
• Highly deviating values (e.g systematic
errors and random errors)
TERMINOLOGIES
DELTA CHECK
• Most ommonly used patient-based QC
technique.
• Requires computerization of test data so
that current results can be compared
with the post-results
INTERFERENCE EXPERIMENTS
• Used to measure systematic errors or
inaccuracy caused by substances other
than the analytes – lipids, biliruubin,
hemoglobin
RECOVERY EXPERIMENTS
• Shows whether a method measures all
the analytes or only part of it (an
estimate of systematic error)
Cuzzamu 2
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT
POINT OF CARE TESTING (POCT)
• Known as Decentralized Testing
• Analytical testing performed outside the
lab, usually by non-laboratorian
personel (e.g blood glucose meter)
PHYSIOLOGIC LIMIT (ADSURD VALUE)
• Helps detect the sample and volume
error, method problem, or incorrect
recording or transmission of results.
ANALYTICAL METHODS
CALORMETRY
SPECTROPHOTOMETRY
• Measurement of light intensity in a
narrower wavelength.
PARTS OF SPECTROPHOTOMETER
LIGHT SOURCE Tungsten lightbulb
ENTRANCE SLIT Prevents stray light or
unwanted into the
monochromator system
MONOCHROMATOR Isolates specific
wavelength of light
EXIT SLIT • Controls the width
of the light
(bandpass)
• Allows a narrow
fraction of light to
reach the
analytical cell
CUVETTE • Also known as
Analytical Cell,
Absorption Cell, or
Sample Cell
Holds the solution whose
concentration is to be
measured
PHOTODETECTOR Detects and converts the
transmitted light into
photoelectric energy
METER/ READ-OUT Galvanometer or ammeter
DEVICE
BEER’S LAW
• States that the concentration of the
unknown substance is directly
CUZZAMU, NIKKA AIRA B [BSMT-4A]
proportional to the absorbed light and
indirectly proportional to transmittance.
DOUBLE-BEAM SPECTROPHOTOMETER
• An instrument that splits
monochromatic light into two
components.
• One beam passes the sample, the other
beam through a reference solution or
blank.
• Additional beam is used to correct
variations in light source intensity
• The absorvance of the sampe can be
read directly
o Double-beam in space
o Double-beam in time
FLAME PHOTOMETRY
• Measures the light emitted by a single
atom burned in a flame.
• Oxygen and acetylene (propane): hot
flame
PRINCIPLE: Excitation of electrons from lower to
higher energy state.
ATOMIC ABSORPTION SPECTROPHOTOMETRY
• Measures the light absorbed by atoms
dissociated by heat.
• Used for detecting small amounts of an
element when concentrations are too
small to be accurately measured by
standard chemical means.
PRINCIPLE: Element is not excited but
dissociated by heat energy from its chemical
bonds and converts the components to atoms.
TURBIDIMETRY
• The amount of light absorbed by a
suspension of particles depends on the
specimen concentration and on the
particle size.
PRINCIPLE: Determines the amount of light
blocked by aparticulate matter suspended in a
turbid solution.
NEPHELOMETRY
• Light scattering depends on the
wavelength and the particle size.
PRINCIPLE: Determines the amount of light
scattered in by a particulae matter suspended in
a turbid solution.
Cuzzamu 3
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT
ELECTROPHORESIS
PRINCIPLE: Migration of charged particles in an
electric field.
A. IONTOPHORESIS – migration of small
charged ions.
B. ZONE ELECTROPHORESIS – migration of
charged macromolecules on
supporting medim.
*Solution more acidic than the isoelectric point =
CATIONS (+) CATHODE (-)
*Solutions more basic than the isoelectric point
= ANIONS (-) ANODE (+)
FACTORS AFFECTING RATE OF MIGRATION
1. Net electric charge of the molecule
2. Size and shape of the molecule
3. Electric field strength (pH and ionic
strength)
4. Nature of the supporting medium
5. Temperature of operation
ISOELECTRIC FOCUSING
• Separating the molecules migrate
through a pH gradient.
• Ideal for proteins with identical sizes but
with different charges.
ADVANTAGES: ability to resolve mixture of
proteins and can detect isoenzymes of CK and
ALP in serm.
DENSITOMETRY
• Reads gel and celluulose acetate
membranes
• Scans and quantitate electrophoretic
patterns.
CHROMATOGRAPHY
PRINCIPLE: Involves the separation of soluble
components in a soltion by specific differences in
physical-chemical characteristics of different
constituents.
2 FORMS OF CHROMATOGRAPHY
PLANAR
• Known as Thin Layer Chromatography
• Involves paper
COLUMN
A. GAS CHROMATOGRAPHY – used for the
separation of steroids, barbiturates,
blood alcohol, and lipids.
CUZZAMU, NIKKA AIRA B [BSMT-4A]
B. LIQUID CHROMATOGRAPHY – used for
fractionation of drug, hormones, lipids,
CHO, and CHONS
FLUOROMETRY
• Determines the amont of ligt emitted by
a molecule after excitation by
electromagnetic radiation.
• Affected by quenching – changes in pH,
temperature, contaminating chemicals,
and photochemical changes
• Uses 2 monochromators:
• Primary – selects wavelengths best
absorbed by the solution
• Secondary – prevents the incident
light from striking the photodetector
ELECTROCHEMISTRY TECHNIQUES
POTENTIOMETRY
• Measurement of differences in voltage
at a constant current.
• Nernst Equation
REFERENCE ELECTRODE – electrode with a
constant voltage.
A. Standard hydrogen
B. Saturated calomel
C. Silver-silver chloride
ION SELECTIVE ELECTRODE
• An electrochemical transducer capable
of responding to one given ion; very
selective.
POLAROGRAPHY
• Measurement of differences in current
at a constant voltage.
• Ilkovic Equation
COULOMETRY
• Measurement of the amount of
electricity at a fixed potential needed to
convert an analyte to adifferent
oxidation state.
• Used to measure chloride ion, CSF, and
sweat samples.
• Faraday’s Law
AMPEROMETRY
• Measurement of the current flow
produced by an oxidation-reaction.
Cuzzamu 4
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
VOLTAMMETRY HORMONES THAT PROMOTES HYPERGLYCEMIA
•Measurement of current after wich a CORTISOL & Decreases intestinal entry
potential is applied to an CORTICOSTEROIDS of glucose into the cell
electrochemical cell. Stress hormone
• Detects very low analyte levels. CATHECOLMINES Inhibits insulin secretion
CARBOHYDRATES GROWTH HORMONE Decreases intestinal entry
• Hydrates of alehyde or ketone (SOMATOTROPHIC) of glucose
derivatives THYROID HORMONES Promotes intestinal
• Presence of double bond and a negative (T3 & T4) absorption of glucose
charge in the enol anion makes glucose Involved in metabolism
an active reducing substance. and hyperthyroidism
GLYCOALDEHYDE Simplest sugar ADENOCORTICOTROPIC Stimulates release of
SUCROSE Most complex sugar HORMONE (ACTH) cortisol
Most common non- SOMATOSTATIN Inhibits and alances insulin
reducing sugar substance and glucagon
(Glucose+Fructose)
FRUCTOSE Commonly seen on fruits DIABETES MELLITUS
(e.g Avocado) • A group of metabollic disorder
characterized by hyperglycemia
MICROALBUMIN TEST resulting from defects in insulin
• Used for kidney function test. secretion, insulin receptors, or both.
• Useful to assist in diagnosis at an early • Also associated as a genetic disorder
stage of glomerular dysfnction and TYPE I DIABETES MELLITUS
before the development stage of • A result of cellular-mediated
proteinuria (>0.5g/day). autoimmune destruction of the B-cells
o <20g/min= NORMAL AER of the pancreas.
o 50-20mg/day = DIABETIC • Known as:
NEPHROPATHY o Insulin-dependent DM
o 20-30mg/g albumin/creatinine o Juvenile Onset DM
ratio = (+) for o Brittle Diabetes
MICROALBMINURIA o Ketosis-Prone Diabetes
• MICROALBUMINURIA: presence of A Cells = Glucagon
microalbumin in urine. B Cells = Insulin
HYPERGLYCEMIA Autoantibodies of B-cells = loss of insulin
• Increase in plasma glucose • Lies on the problem on chromosome 6,
• Diagnosis by glucose tolerance test and HLA DR3 & DR4
postprandial glucose testing. Signs and symptoms
FASTING BLOOD SUGAR (FBS): >126mg/dL • Polyuria
FASTING PLASMA GLUCOSE (FPC): 70-110mg/dL • Polydipsia
GLUCOSE RENAL TRESHOLD: 10-180mg/dL • Polyphagia
• Rapid weight loss
LAB FINDINGS: Increased urine specific gravity • Retinopathy
• Due to glucose molecules TYPE II DIABETES MELLITUS
[macromolecules = heavier in weight] • Has rampant increase in plasma glucose
• Known as:
o Non-Insulin Dependent DM

Cuzzamu 5
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
o Adult/Maturity Onset DM D. Genetic Syndromes
o Stable DM GESTATIONAL DIABETES MELLITUS
o Ketosis-Resistant DM • A disorder characterized by impaired
o Receptor-Deficient DM ability to metabolize CHO, usually
3 MICROVASCULAR PROBLEMS caused by a deficiency of insulin,
• Blockage occuring in pregnancy and disappearing
after delivery.
D Neuropathy The glucose is blocking SCREENING TEST: 1 hour GCT (50g glucose
the interonversion of load) OGTT
substances needed for 24-28 weeks of gestation
the cell ≥140 mg/L plasma glucose = 3hrs GTT
*Tusok-tusok sensation POSSIBLE RESULT
D Nephropathy Kidneys have blood 1st FBS >105 mg/dL
vessels that are thin measurement
which is being blocked 2nd 1 hour <190 mg/dL
by plasma glucose measurement
molecules, interrupting 3rd 2 hour Equal or >165
the blood flow. measurement mg/dL
th
4 3 hour Equal or >145
Rupture of blood vessels measurement mg/dL
lead to KIDNEY FAILURE
D Retinopathy Blood vessel in the eye IMPAIRED FASTING GLUCOSE
was blocked by plasma • Characterized by fasting glucose
glucose molecules that concentrations between normal and
leads to blindness diabetic values
IMPAIRED GLUCOSE TOLERANCE
• The FBS value is <126 but the OGTT is
between normal and diabetic values.
DIABETES INSIPIDUS
• Defeciency of ADH (vassopressin)
released by the posterior gland.
• Characterized by severe polyuria
Increased urine output = Decreased blood
volume = Decreased levels of blood pressure
• Maintains homeostasis

GLUCOSE METHODOLOGIES
OTHER SPECIFIC TYPE OF DM
• Glucose in whole blood is 15% lower
A. Pancreatic Disease
than in serum or plasma
B. Endocrine Disorders
• Serum or plasma must be separated
a. Cushing’s Syndrome
immediately to prevent loss of glucose
b. Pheochromacytoma
(because the cells will consume the
c. Acromegaly
glucose in the blood.)
d. Thyrotoxicosis
• CSF glucose is 60% of the plasma glucose
C. Drug or Chemical Inducers – dilantin &
level (40-0 mg/dL)
pentamidine

Cuzzamu 6
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
HYPOGLYCEMIA Glucose Multiple blood sugar test
• Patient appears healthy Tolerance Test Aids in DM diagnosis
• Low levels of glucose
CHEMICAL METHODS KINDS OF GLUCOSE TOLERANCE TEST
A. Nelson Somogyi – true measure of ORAL GLUCOSE TOLERANCE TEST
glucose • Has two types:
a. Copper Reuction Method o Janney Isaacson Method – single
Glucose Ion + Cuprous Ions + Arsenomolydic Acid dose
= Arsenomolybdenum Blue o Exton Rose Method – divided
b. Folin Wu dose
Glucose Ion + Cuprous Ions + 30 mins 30-60mg/dL above fasting
Phosphomolybdenum Acid 1 hour 20-50mg/dL above fasting
= Phosphomolybdenum Blue 2 hour 5-15mg/dL above fasting
B. Ortho-toluidine – (+) Schiff’s acid base 3 hour Fasting level or below
(green)
ENZYMATIC METHODS INTRAVENOUS GTT
A. Glucose Oxidase (Calorimetric) • Utilized for patients that have
• Specific for B-Dglucose then converts B- gastrointestinal disorders
Dglucose to Gluconic Acid • Done intravenously
0.5g glucose/kg (px body weight) within 3mins
Ex: 65kg = 32.5g glucose
POSITIVE RESULTS FROM NON-DIABETIC PERSONS
Serum/plasma 70-110 mg/dL
5 minutes Max. 250 mg/dL
B. Hexokinase Method 1 hour Significant decrease
• Most specific & reerence method Still >120mg/dL
2 hour <120 mg/dL
3 hour Fasting level

REQUIREMENTS FOR OGTT

1. Patient should be ambulatory
DEXTROSTICS 2. Fasting of 6-8 hours
• Cellular impregnated glucose oxidase, 3. Unrestricted ddiet of 150gms/day for 3
peroxidase, and chromogen days prior to testing
4. The patient should not smoke and drink
• Important for establishing correct insulin
alcohol prior to testing
amount for next dose.
5. Glucose OGTT
SAMPLES FOR ANALYSIS
❖ 70 gms
Random Blood Has no any fasting required
❖ 100 gma
Sugar
❖ 1.75gms glucose/kg BW
Fasting Blood Gives the best indcation of
PROCEDURE FOR OGTT
Sugar (FBS) overall glucose homeostasis
1. Extract FBS (0 hour sample)
2-hour Post Below 120 mg/dL at 2 hours
2. Give glucose load (5-15 mins)
Prandial Blood Evaluates hyperglcemia and
3. Collect 30-min blood sample
Sugar hypoglycemia
4. Collect 1st hour, 2nd hour, 3rd hour blood
samples respectively

Cuzzamu 7
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
RESULTS FRUCTOSAMINE
Urine Negative ➢ Also known as Glycated Albumin:
FBS 95mg/dL (5.2 mmol/L) AMADORI REARRANGEMENT
30 min 150mg/dL (8.3 mmol/L) ➢ Method of monitoring short-term
1 hour 130mg/dL (7.2mmol/L) glucose control (from 2-3 weeks)
2 hour 105 mg/dL (5.8mmol/L) ➢ Useful for monitoring individuals with
3 hour 100mg/dL (5.5 mmol/L) Chronic Hemolytic Anemia and
hemoglobin variants (Hb S or Hb C) –
•FBS – result of fasting process shortened RBC life span
•2-hour – result of insulin and REFERENCE VALUE: 205-285 umol/L
glucagon secretion
THREE POSSIBLE OGTT RESULTS: GLYCOGEN STORAGE DISEASES
1. Normal (Non-diabetic) ➢ Due to deficiency of a specific enzyme
FBS - < 110mg/dL (< 6.1mmol/L) involved in glycogen metabolism.
OGTT – no value exceeding 200mg/dL from the Type I Von-Gierke Gluc-6-PO4
0-hour to the 2nd hour sample Type II Pompe 1,4-Glucosidase
Type IIIa Cori Forbes Debrancher
2. Diabetic Type IV Anderson Brancher
FBS - ≥ 126mg/dL (≥ 7.0mmol/L) Type V Mc Ardle Muscle
OGTT – 2 hour glucose value of ≥ 200mg/dL Phosphorylase
Type VI Hers Liver Phosphorylase
3. Impaired Glucose Tolerance Type VII Tarui Phosphofructokinase
FBS - ≥ 100mg/dL bur < 126mg/dL Type XI Franconi- Gluc-transporter 2
OGTT - ≥ 140mg/dL but < 200 mg/dL Bickel
• Glucose levels do not meet the criteria
for diabetes but are too high to be LIPIDS
consdered normal. ➢ Commonly referred to as FATS; mostly
GLYCOSYLATED HEMOGLOBIN (HbA1C) composed of carbon-hydrogen bonds
• Also called Glycated Hemoglobin ➢ Source of fuel and provides stability to
• Refelcts the average blood glucose level cell membrane
over the previous 2-3 months ➢ Soluble to blood but insoluble to
• Detects the sgars attached to the RBCs inorganic substances
• 2 factors: - Phospholipids
o Average glucose concentration - Cholesterol
o RBC life span - Triglyceride
NORMAL VALUE: 4-6% - Fatty Acid
o 3-6% indicates normal - Fat Soluble
glycosylation Vitamines
o 18-20% indicates prolonged (ADEK)
hyperglycemia PHOSPHOLIPIDS
Sample for Testing : EDTA whole blood (has ➢ Most abndant forms derived from
interference for hemolysis) phosphatidic acid found on the surface
Methods for Testing: Calorimetry, of lipid layers
Electrophoresis, Ion Capture, Immunoassay ➢ Has similar structre with TAG except
Preferred Method: Affinity Chromatography that they have two fatty acids

Cuzzamu 8
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
➢ Amphipathic; conatins polar hydrophilic FREE CHOLESTEROL
head groups and non-polar hydrophobic ➢ 30%
fatty acid side chain. ➢ Serum, plasma, and RBC
➢ Alter fluid surface tension – LUNG
SURFACTANT CHEMICAL REACTIONS
REFERENCE VALUE: 150-380 mg/dL as per IFCC Liebermann End product:
Burchardt Reaction cholestedienylmonosulfonic
KINDS OF PHOSPHOLIPID acid
Lecithin or 70% End color: GREEN
Phosphatidyl Choline Salkowski Reaction End product:
Spingomyelin 20% cholestedienylydisulfonic
Cephalin 10% acid
o Phosphatidyl Ethanolamine End color: RED
o Phosphatidyl Serine
o Iysolecithin + Inositol Phosphatide GENERAL METHODS
*Lecithin-Spingomyelin Ideal Ratio: > 3.5 One-Step Calorimetry
(without respiratory distress syndrome) Method
* Accumulation of Spingomyelin leads to Two-Step Calorimetry + Extraction Method
NIEMANN’S PICK DISEASE Method
CHOLESTEROL Three-Step Calorimetry + Extraction Method
➢ An unsaturated steroid alcohol whose Method + Saponification
transport and excretion is promoted by Four-Step Calorimetry + Extraction Method
estrogen Method + Saponification + Precipitation
➢ Also amphiphatic; can be found on (to be measured on
surface of lipids Spectrophotometer)
➢ Not catabolized by most cells – does not ABELL, LEVY AND BRODIE METHOD
serve as a source of fuel o Current CDC reference method
➢ Has no fasting period required when o Hydrolysis with alcohol, KOH, hexane
testing extraction, and calorimetry with
➢ Plays a big role in production of sex Liebermann Burchardt reagent.
hormones (steroles) TRIGLYCERIDES
DESIRABLE VALUE: < 200 mg/dL ➢ Comprises one molecule of glycerol and
BORDERLINE VALUE: 200-239 mg/dL 3 FA
HIGH VALUE: > 240 mg/dL ➢ Does not contain charged or hydrophilic
CHOLESTEROL ESTER groups ; they are water insoluble and
➢ 70% very hydrophobic
➢ Plasma/Serum ➢ Main storage: ADIPOSE TISSUE
➢ Lecithin Cholesterol Acyl Transferase ➢ Hydrolysis of TAG will yield FA and
(LCAT) converted to energy = excellent insulator
o Catalyzes the esterification of ➢ FASTING REQUIREMENT: 12-14 hours
cholesterol by promoting the REFERENCE VALUES
transfer of fatty acids from Normal < 150mg/dL
lecithin to cholesterol which Borderline 150-199 mg/dL
results in the formation of High
lysolecithn and CE. High TAG 200-499 mg/dL
➢ ApoA1 – activator of LCAT Very High >500 mg/dL

Cuzzamu 9
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]

ATHEROMA – fats floating freely in blood vessels APOLIPOPROTEINS

• Accumulation of fats in blood vessels ➢ Helps keep the lipids in solution during
leads to Hypertension and blood circulation
Atherosclerosis ➢ Interacts with specific cell-surface
receptors
MODIFIED VAN HANDEL AND ZILVERSMITH CHYLOMICRONS
➢ Current CDC reference method ➢ Largest but the least dense (0.95 g/mL)
➢ (+) result: pink-colored product ➢ 90% TAG, 12% CHON
FATTY ACID ➢ Transports exogenous (dietary TAG) to
➢ Linear chain of carbon-hydrogen bonds muscle and depot
➢ Mainly dervied from hydrolysis of TAG in o For energy production
the adipose tissues. ➢ Apo B-48, Apo C and Apo E
➢ Major constituents of TAG and VERY LOW DENSITY LIPOPROTEIN (VLDL)
phospholipids ➢ 65% TAG, 13% Cholesterol, 6-10% CHON
➢ Only small amount is present to plasma, ➢ Transorts endogenous TAG to muscles
most are bound to albumin and fat depot
➢ Important source of energy ➢ Apo B-100, Apo C, and Apo E
LIPOPROTEINS HIGH DENSITY LIPOPROTEIN
➢ Large molecular complexes of lipids with ➢ Smallest but most dense LPP (1.063-
specialized proteins known as 1.21)
apolipoproteins ➢ Transports cholesterol from the tissues
➢ Transport lipids (cholesterol and TAG) to to liver
sites of energy storage and utilization ➢ Vehicle for reverse cholesterol transport
➢ HDL & LDL – transport conjugated ➢ Has high antiheterogenic effect
proteins (must always have the same ➢ 30% Phospholipid, 15 % CE, 45-50%
ratio). CHON
o HDL – aka Bad Cholesterol; free- ➢ Apo A-I, II; Apo C
floating in the body; REFERENCE VALUES:
accumulates cholesterol and LOW: < 40mg/dL
brings it to the liver for HIGH: > 60 mg/dL
metabolism LOW DENSITY LIPOPROTEIN
o LDL – aka Good Cholesterol; low ➢ Most abundant LPP – upto 50% of total
levels of LDL might cause fatty LPP
liver leading to Hepatic Cirrhosis ➢ Major endproduct of VLDL catabolism
and Hepatic Carcinoma ➢ Transport dietary cholesterol to
LIPOPROTEINS AND DIABETES peripheral tissues
• DM is due to increase of sugar (sugar are ➢ Most atherogenic LPP = target of
macromolecules = heavy) therapy
• DM patients develop hypertensive ➢ 50% CE, 18-22% CHON
urgency due to accumulation of sugar ➢ Apo B-100, Apo E
and lipds in the blood vessels (making it REFERENCE VALUES
more constricted + blood flow is Optimal < 100 mg/dL
disturbed). Above 100-129 mg/dL
Optimal
FOAM CELLS – macrophage that engulfs lipids Borderline 130-159 mg/dL

Cuzzamu 10
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
High 10-189 Type 2a RT: (-); clear plasma
Very High 190 or > 190 mg/dL Familial Electrophoresis:
Hypercholesterolemia increased β band
Type 2b RT: (-) ; cloudy plasma
Familial Combined Electrophoresis:
Hyperlipidemia increased pre-β band
and β bands
Type 3 RT:(-); occasionally
Familial cloudy plasma
Dysbetalipoproteinemia Electrophoresis:
“broad β” band =
increased β band
Type 4 RT: (-) ; cloudy plasma –
Familial “fluff” TAG-rich VLDL
DISORDERS ASSOCIATED WITH LIPIDS & LPPS Hypertryglyceridemia particles
DYSBETALIPOPROTEINEMIA Electrophoresis:
➢ Plasma VLDL rich in cholesterol and increased a-2 band
chylomicron remnants Type 5 RT: (+); cloudy plasma
➢ Pesence of E2/2 (rare form of Apo E) Familial Electrophoresis:
➢ VLDL-Choles:TAG = 0.689 Hyperlipoproteinemia increased a-2 band
ABETALIPOPROTEINEMIA (BASSES-KORNZWEIG)
➢ Deficiency of VLDL, LDL, and PROTEINS
Chylomicrons MEDICAL & BIOLOGICAL IMPORTANCE
➢ Defective Apo-B synthesis; defects in ➢ Proteins are involved in:
absorption of vitamins ADEK o Transport of substances in the
➢ Low levels of cholesterol and TAG body
NIEMANN-PICK DISEASE (LIPID STORAGE o Defense function (acts agains
DISEASE) bacterial/viral infection)
➢ Accumulation of spingomyelin in the o Gene expression (Histones
bone marrow, spleen, and lymph nodes controls gene expression and
➢ Deficiency of enzyme spingomyelinase translation)
which is responsible for removal of ➢ Enzymes that catalyzes chemical
phosphorylcholine from spingomyelin reactions in the body are PROTEINS
TANGIER DISEASE ➢ Hormones are proteins; controls many
➢ Markedly decreased almost deficient biochemical events
HDL ➢ Proteins serves as nutrients and also
➢ Increase HDL catabolism involved in storage functions
LIPOPROTEIN LIPASE DEFICIENCY ➢ Acts as buffers
➢ Inability to clear chylomicrons creating ➢ Functions as vitamins
Chylomicronemia Syndrome (TAG: ➢ Can also be infective agents; some toxins
10,000mg/dL) are proteins
➢ Abdominal pain and pancreatitis ➢ Some protein provides structural
FREDERICKSON CLASSIFICATION strength and elasticity system; also
Type 1 Refrigerator Test: (+); components of structural tissues
Familial LPP Deficiency clear plasma ➢ Some proteins have role in contraction
Electrophosis: Normal of muscles

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Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
SIDE NOTES FROM SIR: 8. Proteins acts as buffers because they are
IgG Monomer; small amphoteric substances.
Present in chronic infection CLASSIFICATION OF PROTEINS
IgA Asa labas; secretions (sweat, 1. According to composition or structure
tears, etc.) 2. According to solubility
IgM Malaki (big); Pentamer 3. According to shape
Activats when Malamig (cold) 4. Based on their function; also fond in
Mauuna sa site of infection literature
IgD Di alam CLASSIFICATION BASED ON COMPOSITION
IgE Ellergy (allergy); with Eosinophil ➢ Simple Proteins
▪ Eosinophil is toxic for o Made up of amino acids only
parasites o 1 class only
▪ IgE attaches to parasites o Ex: Human plasma albumin,
to serve as signals for Chymotrypsin, Pepsin, Insulin,
eosinophils when there’s Soyabean, Trypsin inhibitor, and
parasitic infection, tapos Ribonuclease
sasabog yng eosinophil ➢ Conjugated Proteins
undegoing Apoptotic o Proteins containing non-protein
Panel, killing parasites part attached to the protein part
with their toxic CLASSIFICATION BASED ON MEDICAL
eosinophilic granules BIOCHEMISTRY

CHEMICAL NATURE OF PROTEINS

➢ All proteins have polymers of amino
acids. The amino acids in proteins are
united through “Peptide” linkage.
Sometimes, proteins are also called
‘Polypetides’ because they contain many
peptide bonds.
PROPERTIES OF PROTEINS
1. High molecular weight
2. Colloidal in nature
3. Large particle size BASED ON ACTS (REVIEW CENTER):
4. Different kinds of proteins are soluble to 1. Glycoproteins – 10-40% CHO
different kinds of solvents (haptoglobin)
5. Proteins differ in shape 2. Mucoproteins - > 40% CHO (mucin)
6. Some proteins yeilds amino acids only 3. Lipoproteins
on hydrolysis; others produce amino 4. Metalloproteins – ex: Seruloplasmin
acids plus other types of molecules 5. Nucleoproteins
7. Charge properties
a. Charge of a protei depends on ➢ Derived proteins
the surroundings like amino o Proteins formed from simple
acids. By changing of the pH of and conjugated proteins
surroundings, the charge of Primary Formed from natural proteins by
protein can be altered. Derived action of heat or alcohol. (Ex:
Proteins Cooked-egg albumin

Cuzzamu 12
Clinical Chemistry
Lecture / PPT based
Professor: Angelo Del Rosario, RMT CUZZAMU, NIKKA AIRA B [BSMT-4A]
Secondary Formed from partial hydrolysis of PLASMA PROTEINS
Derived proteins (Ex: Peptone, gelatin,
Proteins peptides)

CLASSIFICATION ACCORDING TO SOLUBILITY

Albumins Soluble in water and salt
solutions
Ex: Albumin of plasma, egg
albumin and lactalbumin of milk
Globulins Sparingly solble in water but
soluble in salt solutions
Ex: Globulins of plasma,
ovaglobulins of egg,
ALBUMIN
lactoglobulin of milk
➢ Composed of 585 amino acid
Glutelins Soluble in dilute acids and
➢ Present in the highest concentration in
alkalies
serum
Ex: Glutenin of wheat, oryzenin
➢ Synthesized in the liver
of rice, zein of maize
➢ Genral transport protein (binds various
Protamins Soluble in ammonia and water
subsances in the blood)
Ex: Salmine from Salmon
➢ Maintains osmotic pressure and
Histones Soluble in water and dilute acids
indicator of nutritional status
Ex: Histones present in
➢ A circulating reservoir of amino acids
chromatin
➢ Sensitive and highly prognostic marker
Prolamines Soluble in dilute alcohol and in cases of cystic fibrosis
insoluble in water and alcohol ➢ A negatie ACUTE PHASE REACTANT
Ex: Giladin of wheat, zein of corn o Lowest plasma levels are seen in
Sclero Insoluble in water and dilute active nephrotic syndrome
Proteins acids and alkalies o REFERENCE VALUE: 3.5-5.0 g/L
Ex: Collagen, Elastin, and Keratin Analbuminemia Absence of albumin
Bisalbuminemia Presence of 2 bands in the
albumin region
CLASSIFICATION BASED ON SHAPE Hypoalbuminemia Decreased levels of albumin,
➢ Globlar Proteins – polypeptide chains of typical of nephritic syndrome
these protens are folded into compact
globular (spherical) shape.
DECREASED INCREASED
o Symmetrical
Liver Disorders Dehydration
o Ex: Hemoglobin, Myoglobin,
Gastrointestinal Excessive albumin
Albumin, Lysozome,
Disease administration
Chymotrypsin
Malabsorption
➢ Fibrous Proteins – polypeptide chains
Muscle Waisting
are extended along one axis.
Disease
o Elongated & Asymmetrical
o Ex: a-Keratin, B-Keratin, Severe Burns
Collagen, Troponin, and Elastin Renal Diseases

Cuzzamu 13