CC DAY 1 Flashcards
ISO means
International Organization for Standardization
IUPAC means
International Union of Pure and Applied Chemistry
CLSI means
Clinical Laboratory Standards Institute
SAMHSA means
Substance Abuse and Mental Health Services Administration
this certifies labs to conduct forensic drug testing for federal agencies
SAMHSA
Blanking technique composition
serum w/ rgt
For correcting absorbance caused by reagent’s color and Hb
Blanking technique
used to subtract intrinsic absorbance of the sample usually caused by hemolysis, icterus, turbidity, or drug interference
Sample blank
FEP colors:
Sodium
yellow
FEP colors:
Copper
Blue
FEP colors:
Magnesium
Blue
FEP colors:
Lithium
red
FEP colors:
Rubidium
red
FEP colors:
Potassium
Purple
AAS
most common burner
pre-mix long-path burner
Flameless AAS use _____ instead of flame
electricity
AAS
used to increase sensitivity for atomic emission
Inductively coupled plasma (ICP)
In fluorometry/molecular luminescence spectrophotometry, which part controls light intensity?
attenuator
In fluorometry/molecular luminescence spectrophotometry, which part selects wavelength that is best absorbed by the solution?
Primary filter
In fluorometry/molecular luminescence spectrophotometry, which part detects fluorescing sample?
Detector
Main disadvantage of fluorometry
Quenching
- should not be acidic
- should not be hot
- interference of some chemicals
These are the most frequently used sources of excitation radiant energy in flurometry
Gas discharge lamps (mercury and xenon)
Most spectrofluorometers use a high-pressure _______ lamp
xenon
How many times is fluorometer more sensitive than spectrophotometer?
1000x
Fluorescence _________ is widely used for the detection of therapeutic and abused drugs
Fluorescence polarization
emission of light is created from a chemical or electrochemical rxn
chemiluminescence
No excitation, no monochromator
chemiluminescence
measures light blocked by a particle in a solution
turbidimetry
turbidimetry is dependent on what factors?
Concentration
Size
determines the amount of scattered light
nephelometry
T/F:
Nephelometry is more sensitive than turbidimetry
TRUE
Turbidimetry photodetectors: 1
Neph photodetectors: 2
Where are the photodetectors located in nephelometry
90 degrees
Forward angle
In nephelometry, if macromolecules > wavelength, which angle is measured?
Forward
Nephelometry depends on what factors
size
wavelength
Coulter counter is application of what type of technique?
Laser light
narrow spectral width and small cross-sectional area with low divergence
laser light
for determination of structure and ID of samples
laser light
a molecule, such as protein, whose net charge can be either positive or neg
ampholyte
pH where protein has no net charge
Isoelectric point
At what pH do proteins migrate from cathode to anode?
8.6
Fastest protein to migrate to anode
ALBUMIN
a1
a2
b
gamma
type of electrophoresis involving small ions
Iontophoresis
migration of charged macro molecules
zone electrophoresis
Type of electrophoresis where separation is performed in narrow-bore fused silica capillaries and molecules are separated by EOF
Capillary Electrophoresis
In capillary electrophoresis, molecules are separated by?
Electro-osmotic flow (EOF)
In capillary electrophoresis (+/-) charge moves faster
Positive
(moves towards the negative charge)
(negative charges are attracted to the walls of silica and moves more slowly)
Involves molecules migrating through a pH gradient; ideal for separating proteins of identical size but different net charges
Isoelectric focusing
separate proteins into as many as 12 zones
High-resolution protein electrophoresis
Component of electrophoresis that serves as the driving force
electricity
Component of electrophoresis that separates by charge and molecular size
supporting medium - starch gel
Component of electrophoresis that separates by molecular size and is used in isoelectric focusing
Cellulose acetate
Component of electrophoresis that is neutral and does not bind to protein and separates electrical charge
agarose gel
Component of electrophoresis that is neutral and separates by charge and size; for isoenzymes
supporting medium - polyacrylamide gel
What methods are used for detection and quantitation?
Densitometry
UV
measures absorbance of stain; scan and quantitate electrophoretic pattern
Densitometry
In electrophoresis, hemoglobin hemolysate (must/must not) be concentrated
must not
** Urine is concentrated
** Serum with high protein conc. is diluted
Stain used for electrophoresis of CSF proteins
silver nitrate
Stain used for electrophoresis of lipoprotein
Fat red 7B
Oil Red O
Stain used for electrophoresis of LD isoenzymes
nitrotetrazolium blue (NTB)
separation of soluble components in a solution
chromatography
distance leading edge of component moves/total distance of solvent
Retention factor value
gold std for drug testing
GC-MS
detect 20 inborn errors of metabolism (newborn screening)
MS/MS (tandem mass spectrometry
HPLC uses _________ for fast separations
pressure
rapid Hba1c and hg disease is detected using what technique
HPLC
uses sugar and amino acid
paper chromatog (whatman paper)
unknown sample + known sample + indicator
volumetric
schales and schales method detects which anion
chloride
(volumetric)
measurement of osmolality
osmometry
inversely proportional to osmolality
Freezing point
Vapor pressure
directly proportional to osmolality
Boiling point
Osmotic pressure
most commonly used method for measuring the changes in colligative properties in a solution
Freezing point depression osmometry
Reference solution for osmometers
NaCl
For each osmole, freezing pt is lowered by
-1.86 C
For each osmole, vapor pressure is lowered by
0.3 mmHg/Torr
For each osmole, boiling point is raised by
0.52 C
measured osmolality - calculated osmolality
Osmolal gap
Indirectly indicates the presence of osmotically active substances
Osmolal gap
measurement of current or voltage generated by the activity of a specific ion
electrochemistry
a type of potentiometric, ion-selective electrode, used to separate membrane from sample solution
ISE membrane
ISE membranes:
glass aluminum silicate is used for measurement of
sodium
ISE membranes:
antibiotic valinomycin gel is used for measurement of
K+
ISE membranes:
diocytophenyl phosphonate is used for measurement of
Calcium
ISE membranes:
nonactin and monactin is used for measurement of
NH4+
measures the amount of current produced through the oxidation or reduction of the substance to be measured at an electrode held at a fixed potential
Amperometry
electrode used for measurement of pO2 (amperometry)
clarke
measures potential difference between an indicator electrode and a reference electrode
potentiometry
reference electrodes in potentiometry
Calomel
Ag - Ag Cl
electrode for pCO2 (potentiometry)
severinghaus
analyte which uses nernst equation (potentiometry)
pH
Coulemetry is based on which law
Faraday’s
number of equivalent weights of a reactant oxidized or reduced is directly proportional to the quantity of electricity used in the reaction
Coulometry
voltage at which sharp rise in current occurs characteristic of the electrochemical rxn involved
Polarography
measures the differences in current at a constant voltage
Polarography
amount of inc in current is proportional to the conc. of analyte
Polarography
trace metal ions in the solution is reduced and plated onto anodic electrode and is mainly used for analysis of lead
Anodic stripping voltammetry
