Laboratory Safety, Basic Units & Conversion Factors Flashcards
It is a policies that mandate measures to reduce or eliminate exposure to hazard
Work Practice Control
-Hand washing after each patient contact -Cleaning surfaces with disinfectants
-Avoiding unnecessary use of needles and sharps and not recapping
-Red bag waste disposal
-Immunization for hepatitis
-Job rotation to minimize repetitive tasks -Orientation, training, and continuing education -No eating, drinking, or smoking in laboratory
-Signage/warning signage
Work Practice Control
A safety features built into the overall design of a product
Engineering controls
-Puncture-resistant containers for disposal and transport of needles and sharps
-Safety needles that automatically retract after removal
-Biohazard bags
-Splash guards
-Volatile liquid carriers
-Centrifuge safety buckets
-Biological safety cabinets and fume hoods -Mechanical pipetting devices
-Computer wrist/arm pads
-Sensor-controlled sinks or foot/knee/ elbow controlled faucets
Engineering controls
A barriers that physically separate the user from a hazard
PPE
-Nonlatex gloves Isolation gowns
-Masks, including particulate respirators
-Face shields
-Protective eyewear (goggles, safety glasses)
PPE
-Chemical-resistant gloves;
-subzero (freezer) gloves; thermal gloves
-Hearing protection (earplugs or earmuffs) -Eyewash station
-Safety shower
-Fire extinguisher
-Laboratory spill kit
-First aid kit
Emergency equipment
Thermodynamic temperature
Kelvin
Celsius temperature
Degree celsius
A predominant practice for temperature measurement uses:
Celsius or centigrade
The SI designation for temperature:
Kelvin
Unit of enzymes. A catalytic activity.
Katal
Formula of Celsius to Fahrenheit
°C (9/5) + 32
Formula of Fahrenheit to Celsius:
(°F - 32) 5/9
Exa
10’18
Peta
10’15
Tera
10’12
Giga
10’9
Mega
10’6
Kilo
10’3
Hecto
10’2
Deka
10’1
Deci
10’-1
Centi
10’-2
Milli
10’-3
Micro
10’-6
Nano
10’-9
Pico
10’-12
Femto
10’-15
Atto
10’-18
Conversion factor of Albumin:
10
Conversion factor of Bilirubin
17.1
Conversion factor of BUN
0.357
Conversion factor of Na, K, Cl
1
Conversion factor of Cholesterol
0.026
Conversion factor of Creatinine
88.4
Conversion factor of Glucose
0.0555
Conversion factor of Thyroxine
12.87
Conversion factor of Total Protein
10
Conversion factor of Triglycerides
0.0113
Conversion factor of Uric Acid
0.0595
Wavelength of visible
400-700nm
Wavelength of UV region
<400nm
Wavelength of Infrared region
> 700 nm
Frequency is inversely proportional to wavelength
Planck
A measurement of light transmitted by a solution
Spectrophotometry
Polychromatic light ; many color
Light/Radiant source
Most commonly used light
Tungsten light bulb/incandescent tungsten/tungsten iodide lamp
Light source used in AAS
Hollow cathode lamp
Most commonly used light in UV
deuterium discharge lamp and the mercury arc lamp
Wavelength of hydrogen deuterium lamp
200-375 nm
It minimizes unwanted stray light.
Entrance slit
The most common cause of loss of linearity
Stray light
Major effect of stray light
Absorbance error
It detects stray light
Cutoff filters
It verifies absorbance accuracy on linearity
Neutral filter & dichromate solution
Increase concentration, increase absorbance
Linearity
It isolates specific wavelengths of light.
Monochromator
Most commonly used monochromator
Diffraction gratings
Short wavelengths are refracted more than long wavelengths
Prism
A least expensive, not precise, they are simple, inexpensive, and useful monochromator
Colored glass filter
Check wavelength accuracy
Didymium or holmium oxide filter
Mercury arc lamp
Measurement of assay at two different wavelength
Bichromatic analysis
It controls the bandpass or band width
Exit slit
It holds the solution
Cuvet
Also called absorption cell/analytical cell/sample cell
Cuvet
Examples of cuvet
Etched
Square cuvette
Scratched optical surface
Alumina silica
Glass cuvette
Quartz
Most commonly used cuvet
Alumina silica
Cuvet with less error from lens effect
Square cuvette
Cuvet that scatters light
Scratched optical surfaces
Cuvet for visible range but absorbs UV
Glass cuvettes
Cuvet for UV radiation
Quartz
Low absorbance is equal to ____ transmittance
High
High absorbance is equal to ____ transmittance
Low
Unknown substance is directly proportional to absorbed light and inversely proportional to transmitted light
Beer’s law
High concentration is equal to ____ absorbance
High
The absorbance increases exponentially with an increase in the light path
Lambert law
It detects and converts transmitted light to electrical energy
Photodectector
The most common type of photodetector and has an excellent sensitivity
Photomultiplier tube
It should never be exposed to room light because it will burn out
Photomultiplier tube
Simplest detector. It requires no external voltage source
Barrier cell/Photocell/Photovoltaic Cell
It differs in that an outside voltage is required for operation
Phototube
Respond to a specific wavelength UV/visible
Photodiode
Designed with _________photodiodes that are arranged in linear fashion
256-2048
Displays output result
Meter or readout device
Simplest type of beam. One measurement at a time.
Single beam spectrophotometer
Permit automatic correction of sample and reference absorbance
Double Beam spectrophotometer
Use two photodetectors
Double Beam in space
Use one photodetector
Double Beam in time
A solution consisting of all the components of a reaction except the analyte
Blank
It has no sample.
Contains the same reagents used for the test.
Adjust the spectrophotometer to 100% transmittance.
No correction for interfering chromogen or lipemia.
Reagent blank
Remedy in blanking technique.
Ultracentrifuge
Serum with Reagent. For correcting absorbance caused by reagents color and hemoglobin.
Turbidity by lipid is not corrected.
Blanking technique
It removes turbidity
Ultracentrifuge
It is used to subtract the intrinsic absorbance of the sample usually caused by hemolysis, icterus, turbidity, or drug interference
Sample blank
It has its first Reagent.
It has no Reagent for color development.
Sample blank
Measure the light emitted by a single atom burned in a flame.
Flame emission photometry
Flame color from hottest to least hot
Blue
White
Yellow
Orange
Red
Hottest flame color
Blue
Flame color of sodium
Yellow
Flame color of potassium
Violet
Flame color of lithium/rubidium
Red
Flame color of magnesium/copper
Blue
Internal standard in FEP
Lithium and Cesium
Doesn’t create color reaction
Unexcited molecule
No color reaction
Dissociation
Light intensity of atoms that are emitting energy
Concentration
Measures light absorbed by atoms dissociated by heat
Atomic Absorption Spectrophotometry
Bring the metal analyte from molecular form into its Atomic form at ground state.
It accepts the sample/cuvet
Flame
Unexcited Trace Metals
“CaCoMa LeAlLi Zinc”
Calcium
Copper
Magnesium
Lead
Aluminum
Lithium
Zinc
Most common burner
Premix long path burner
Uses electricity to break the chemical bonds instead of flame.
Flameless AAS
Used in increased sensitivity for atomic emission
Inductively Coupled Plasma
Periodic table of elements assay
ICP + MS
Controls light intensity
Attenuator
Selects wavelength that is best absorbed by the solution
Primary filter
Detects fluorescing sample
Detector
Most frequently used sources of excitation radiant energy
Gas discharge lamp (mercury & xenon arc)
Most spectrofluorometers use a:
High pressure xenon lamp
Measure amount of light intensity emitted by a molecule after excitation by electromagnetic radiation
Fluorometry/Molecular Luminescence Spectrophotometry
______ more sensitive than spectrophotometer
1000x
Disadvantage of fluorometry
Quenching effect
Temperature in fluorometry should not be:
High temperature
pH in fluorometry should not be:
Acidic
Widely used for the detection of therapeutic and abused drugs
Fluorescence Polarization
Enhance of chemiluminescence
Enzyme (Bioluminescence)
It is more sensitive than fluorometry
Chemiluminescence
Emission of light is created from a chemical or electrochemical reaction. No excitation, no monochromator.
Chemiluminescence
Reducing agents are:
Donors
Oxidizing agents are:
Acceptor
It speeds up chemical reactions.
Enzyme
Light blocked by a particle in a solution
Turbidimetry
Turbidimetry is dependent on:
Concentration and Size
For measuring abundant large particles
Turbidimetry
It determines the amount of scattered light
Nephelometry
It is more sensitive than turbidimetry
Nephelometry
For measuring antigen-antibody complexes
Nephelometry
Nephelometry depends on:
Wavelength and particle size
Macromolecules > Wavelength measures the:
Forward angle
Application of laser lights
Coulter counter
More sensitive than spectrophotometer
Laser lights
Narrow spectral width and small crossed sectional area with low divergence. The determination of structure and identification of samples.
Laser lights
pH where protein has no net charge
Isoelectric point
A molecule, such as protein, whose net charge can be either positive or negative due to amino acid.
Ampholyte
What pH does proteins migrate and divide
pH 8.6
Migration of small ions
Iontophoresis
Electrophoresis of proteins from fastest to slowest:
Albumin
A1 globulin
A2 globulin
Beta globulin
Gamma globulin
Migration of charged macromolecules
Zone electrophoresis
Migration of charged particles in electrophoresis
Towards anode
In capillary electrophoresis, molecules are separated by:
Electro osmotic flow
In capillary electrophoresis, a charge that moves faster
Positive charge
In capillary electrophoresis, a charge that moves slower
Negative charge
A separation is performed in narrow bore fused silica capillaries
Capillary electrophoresis
Separate proteins into as many as 12 zones
High resolution protein electrophoresis
Ideal for separating proteins of identical size but with different net charge
Isoelectric focusing
In isoelectric focusing, molecules migrate thru a:
pH gradient
Components of electrophoresis
Driving force
Support medium
Buffer
Sample
Detecting system
Separates the charge and molecular size
Starch gel
Separates by molecular size used in isoelectric focusing
Cellulose acetate
Neutral, does not bind to protein and separates by electrical charge
Agarose gel
Neutral, separates by charge and size, separates protein into 20 fractions. It is for isoenzymes
Polyacrylamide gel
Measures absorbance of stain. Scan and quantitate electrophoretic pattern.
Densitometry
Specimen for high protein concentration and must be diluted
Serum
Specimen for low protein and concentrated
Urine and CSF
Specimen for no concentration
Hemoglobin hemolysate
Used for CSF proteins
Silver nitrate
Used for lipoprotein
Fat red 7b and oil red O
Used for lactate dehydrogenase isoenzymes.
Nitrotetrazolium blue
Separation of soluble components in a solution
Chromatography
Distance leading edge of component moves/total distance of solvent
Retention Factor (RF) Factor
Chromatography for sugar and amino acid
Paper chromatography (Whatman paper)
Chromatography for drug screening
Thin layer chromatography (silica gel or alumina)
Gold standard for drug testing
GC-MS
How many days should the test be challenged after receipt of the result through GC-MS
15 days
Detects 20 inborn errors of metabolism
MS/MS Tandem Mass spectrometry
Most widely used. It uses pressure for fast separations.
HPLC
Uses of HPLC
Rapid HbA1c and Hb disease
Test use in volumetric
Scales and scales method (Chloride test)
Volumetric formula
Unknown sample + known sample + indicator
Measurement of osmolality or concentration
Osmometry
Osmotic particles
Glucose
Urea nitrogen
Sodium
Ethanol
Alcohol
Colligative properties of solution
Osmotic pressure
Boiling point
Freezing point
Vapor pressure
Most commonly used method for measuring the changes in colligative properties in a solution.
Freezing point
Reference solution in osmometry
Sodium chloride
How many degrees does the freezing point be lowered by:
-1.86 °C
Vapor pressure is lowered by:
0.3 mmHg or Torr
How many degrees does the boiling point raised by:
0.52 °C
Indirectly indicates the presence of osmotically active substances
Osmolal gap
The difference between the measured osmolality and the calculated osmolality
Osmolal gap
Formula of osmolal gap
Measured osmolality - calculated osmolality
True or false:
FaVor is inversely proportional to osmolality
True
True or false:
FaVor increases while osmotic pressure decreases
False
*FaVor decreases while osmotic pressure increases
True or false:
BP & OP is inversely proportional to osmolality
False
*BP & OP is directly proportional to osmolality
True or false:
BP & OP increases, osmolality increases
True
Measurement of current or voltage generated by activity of specific ion
Electrochemistry techniques
A type of potentiometric, ion-selective electrode, used to separate membrane from sample solution
ISE Membrane
NH4+ analysis
No action & monactin
Electrolyte in glass aluminum silicate
Sodium
Electrolyte in antibiotic valinomycin gel
Potassium
Electrolyte in dioctylphenyl phosphate
Calcium
Test for pO2
Clarke electrode
Test for glucose
Glucose oxidase
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
Uses in potentiometry
pH and pCO2
Reference electrode of potentiometry
Calomel or Silver-silver chloride
Electrode for pCO2
Severinghaus
Equation use for pH
Nernst equation
Use in coulometry
Chloride coulometer
It is based on Faraday’s law.
A number of equivalent weights of a reactant oxidized or reduced is directly proportional to the quantity of electricity used in the reaction
Coulometry
It is based on polarography.
Trace metal ions in the solution are reduced and plated onto anodic electrodes.
Metal stripped off anode.
For the analysis of lead
Anodic stripping voltammetry
Voltage at which sharp rise in current occurs characteristic of the electrochemical reaction involved
Polarography
It is the measurement of difference in current at a constant voltage
Polarography
