ANALYTICAL METHODS Flashcards
(147 cards)
1
Q
IT IS TRANSMITTED VIA ELECTRONIC WAVES BY FREQUENCY AND WAVELENGTH
A
ENERGY
2
Q
RELATIONSHIP BETWEEN WAVELENGTH AND ENERGY
A
PLANCK’S FORMULA
3
Q
DISTANCE BETWEEN TWO SUCCESSIVE PEAKS EXPRESSED IN NANOMETERS
A
WAVELENGTH
4
Q
400-470NM
A
VISIBLE SPECTRUM
5
Q
<400NM
A
UV LIGHT
6
Q
> 700NM
A
INFRARED REGION
7
Q
IT IS THE NUMBER OF VARIATIONS OF WAVE MOTION PER SECOND
A
FREQUENCY
8
Q
RELATIONSHIP OF FREQUENCY AND WAVELENGTH
A
INVERSELY PROPORTIONAL
9
Q
MEASUREMENT OF LIGHT IN A NARROWER WAVELENGTH
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SPECTROPHOTOMETRIC MEASUREMENT
10
Q
MEASUREMENT OF LIGHT WITHOUT WAVELENGTH
A
PHOTOMETRIC MEASUREMENT
11
Q
MEASURES LIGHT TRANSMITTED TO DETERMINE ITS CONCENTRATION
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SPECTROPHOTOMETRY
12
Q
EMITS RADIATION TAHT CHANGES IN INTENSITY
A
CONTIMUUM SOURCE
13
Q
EMITS LIMITED RADIATION AND WAVELNGTH
A
LINE SOURCE
14
Q
INTENSE BEAM LIGHT IS DIRECTED THROUGH
A
MONOCHROMATOR AND SAMPLE
15
Q
COMMONLY USED LIGHT SOURCE IN VISIBLE AND IR
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TUNGSTEN LIGHT BULB
16
Q
IT GIVES ACCURATE ABSORBANCE MEASUREMENTS AND RESPOSNSE TO CHANGE IN LIGHT INTENSITY
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LINEARITY
17
Q
FACTORS FOR CHOOSING LIGHT SOURCE
A
- SPECTRUM
- STABILITY
- SOURCE
- RANGE
- TEMPERATURE
18
Q
ALTERNATIVES FOR COLORIMETRY UV LIGHT
A
MERCURY ARC
DEUTERIUM LAMP
HYDROGEN LAMP
XENON LAMP
19
Q
IT MINIMIZES UNWANTED STRAY LIGHT
A
ENTRANCE SLIT
20
Q
IT IS A WAVELENGTH OUTSIDE THE BAND CAUSES ABSORBANCE ERROR
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STARY LIGHT
21
Q
MOST COMMON CAUSE OF LOSS OF LINEARUTY AT HIGH ANALYTE CONCENTRATION
A
STRAY LIGHT
22
Q
KINDS OF MONOCHROMATORS
A
PRISMS
DIFFRACTION GRATINGS
FILTER
23
Q
- GLASS, QUARTZ, SODIUM CHLORIDE
- REFRACTED TO MORE DENSE GLASS
- CAN BE ROTATED
A
PRISMS
24
Q
MOST COMMONLY SED AND BETTER RESOLUTION THAN PRISM
A
DIFFRACTION GRATINGS
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--CROWN GLASS
- BENT TO SHARP CORNER
DIFFRACTION GRATINGS
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FILTERS HAS SEMI TRANSPARENT SILVER FILMS SUCH AS
MAGNESIUM FLUORIDE
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LIGHT BASED ON CONSTRUCTIVE INTERFERENCE OF WAVES
FILTERS
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- REFLECTED
- PASS A WIDE BAND OF RADIANT ENERGY
- LOW TRANSMITTANCE
FILTERS
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CONTROLS WIDTH OF LIGHT BEAM
ALLOWS ONLY NARROW FRACTION OF SPECTRUM
EXIT SLIT
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THE SPECTRAL PURITY OF SPECTROPHOTOMETER IS REFLECTED BY
BANDPASS
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REQUIRED BANDPASS FOR ACCURATE ABSORBANCE MEASUREMENT
<1/5
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TEH DEGREE OF WAVELENGTH ISOLATION DEPENDS ON
DEVICE
ENTRANCE SLIT
EXIT SLIT
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THE RANGE PF WAVELENHTHS BETWEEN POINTS
1/2 PEAK TRANSMITTANCE
BANDPASS
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MOST COMMONLY USED CUVETS
ALUMINA SILICA GLASS
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CUVET FOR MEASUREMENT OF SOLUTION REQUIRES UV SPECTRUM
QUARTZ/PLASTIC
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CUVETS TRANSMIT LIGHT AT >220NM
SILICA OR SOFT GLASS
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SOLUTIONS THAT PRODUCES ETCHING
ALKALI SOLUTIONS
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DETECTS AND CONVERTS TRANSMITTED LIGHT INTO PHOTOELECTRIC ENERGY
PHOTODETECTOR
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- SIMPLIEST, CHEAP, TEMPERATURE SENSITIVE
- SELENIUM PLATE WITH SILVER COVER
- NO EXTERNAL VOLTAGE
BARRIER LAYER CELL/ PHOTO CELL/PHOTOVOLTAIC CELL
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IT IS USED IN FILTER PHOTOMETERS WITH WIDE BANDPASS
BARRIER CELL/PHOTOCELL/PHOTOVOLTAIC CELL
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- CATHODE AND ANODE
- GLASS CASE
- PHOTOSENSITIVE MATERIAL; GIVES OFF ELECTRONS
- REQUIRES EXTERNAL VOLTAGE
PHOTOTUBE
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MOST COMMON TYPE OF DETECTORS
PHOTOMULTIPLIER TUBE
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- MEASURES VISIBLE AND UV REGIONS
- EXCELLENT SENSITIVITY AND RAPID
- DETECTS AND AMPLIFIES RADIANT ENERGY
PHOTOMULTIPLIER TUBE
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DISADVANTAGE OF PM TUBE
WILL BURN OUT AT ROOM LIGHT
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- EXCELLENT LINEARITY
- MULTITUDE OF WAVELENGTHS
- DETECTS LESS AMOUNT OF LIGHT
PHOTODIODE
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DISPLAYS OUTPUT OF DETECTION SYSTEM
GALVANOMETER/AMMETER
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IT STATES THAT CONCENTRATION OF UNKNOWN SUBSTANCE IS DIRECTLY PROPORTIONAL TO ABSORBED LIGHT AND INVERSELY PROPORTIONAL TO TRANSMITTED LIGHT
BEER'S LAW
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IS PROPORTIONAL TO THE INVERSE LOG OF TRANSMITTANCE
ABSORBANCE
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IT IS THE RATIO OF RADIANT ENERGY TRANSMITTED OVER INCIDENT RADIANT ENERGY
PERCENT TRANSMITTANCE
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INSTRUMENT THAT SPLITS LIGHT INTO 2 COMPONENTS
DOUBLE BEAM SPECTROPHOTOMETER
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PASSAGE WAY OF BEAM ACCORDING TO DOUBLE BEAM SPECTROPHOTOMETER
THROUGH THE SAMPLE AND REFERENCE SOLUTION OR BLANK SOLUTION
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WHAT DO ADDITIONAL BEAM CORRECTS
LIGHT SOURCE INTENSITY
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TYPE OF SAMPLE BEAM
- USES 2 PHOTODETECTORS
- FOR SAMPLE AND REFERENCE BEAM
DOUBLE BEAM IN SPACE
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TYPE OF DOUBLE BEAM
- 1 PHOTODETECTOR
- ALTERNATELY ON MONOCHROMATIC LIGHT THROUGH SAMPLE AND REFERENCE CUVET
- CHOPPER
DOUBLE BEAM IN TIME
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THE OBSERVED COLOR FROM LIGHT ABSORBED FROM SOLUTION
COMPLEMENTARY COLOR
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THEY ARE GREATER IN BLUE REGION THAN IN RED REGION
TURBIDITY
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USED TO CHECK WAVELENGTH ACCURACY
DIDYMIUM OR HOLMIUM OXIDE FILTER
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VERIFY ABSORBANCE ACCURACY ON LINEARITY
NEUTRAL DENSITY FILTERS AND DICHROMATE SOLUTION
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CONTAINS SERUM WITHOUT REAGENT
BLANKING TECHNIQUE
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CORRECTS ABSORBANCE CAUSED BY COLOR OF REAGENTS
REAGENT BLANK
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MEASURES ABSORBANCE OF SAMPLE IN THE REAGENT IN ABSENCE OF ENDPRODUCT
CORRECTS OPTICAL INTERFERENCE
SAMPLE BLANK
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IT MUST BE KEPT CONSTANT TO HAVE ABSORBANCE PROPORTIONAL CONCENTRATION
LIGHT PATH
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AMOUNT OF LIGHT ABSORBED MAY VARY ACCORDING TO
CONCENTRATION, PH OR TEMPERATURE
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MEASURES: LIGHT EMITTED BURNED IN FLAME
PRINCIPLE: EXCITATION
LIGHT SOURCE: FLAME
FLAME EMISSION PHOTOMETRY
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INTERNAL STTANDARD OF FEP THAT IS USED TO CORRECT FOR VARIATIONS IN FLAME AND ATOMIZER CHARACTERISTICS
LITHIUM/ CESIUM
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IT IS MEASURED BY FEP
EXCITED IONS (SODIUM AND POTASSIUM)
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MEASURES: LIGHT ABSORBED BY HEAT
LIGHT SOURCE: HOLLOW CATHODE TUBE
ACCURATE, PRECISE, VERY SPECIFIC
NO INTERNAL STANDARD
AAS
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IT IS USED TO MODULTAE THE LIGHT SOURCE
CHOPPER
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IT IS ADDED TO AVOID CALCIUM INTERFERENCE
LANTHANUM OR STRONTIUM CHLORIDE
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INTERFERES AAS
CHEMICAL
MATRIX
IONIZATION
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PRINCIPLE: UNKOWN SAMPLE REACTS WITH KNOWN SOLUTION WITH INDICATOR
VOLUMETRIC/ TITRIMETRIC
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EXAMPLES ARE CHLORIDE AND CALCIUM TEST
VOLUMTERIC/ TITRIMETRIC
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FOR LARGE PARTICLES AND BACTERIAL SUSPENSION
PRINCIPLE: LIGHT BLOCKED
DEPENDS ON: CONCENTRATION AND SIZE
MEASURES: REDUCTION OF LIGHT
TURBIDIMETRY
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MEASURES: ANTIGEN ANTIBODY COMPLEXES (PROTEIN)
PRINCIPLE: SCATTERED LIGHT
DEPENDS ON: WAVELENGTH AND SIZE
PM TUBE
NEPHELOMETRY
75
IT DETERMINES THE NET CHARGE ON A PROTEIN
ACIDIC AND BASIC AMINO ACIDS
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DURING ELECTROPHORESIS PROTEINS ARE?
NEGATIVELY CHARGE
| MOVES IN ANODE
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IT IS THE MIGRATIONOF SMALL CHARGED IONS
IONTOPHORESIS
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MIGRATION OF CHARGED MACROMOLECULES
ZONE ELECTROPHORESIS
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NET CHARGE EITHER POSITIVE OR NEGATIVE DEPENDING ON PH
AMPHOTERIC
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MOVEMENT OF BUFFER IONS AND SOLVENT RELATIVE TO FIXED SUPPORT
ELECTROENDOSMOSIS
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FACTORS AFFECTING RATE OF MIGRATION
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ELECTRIC CHARGE
SIZE AND SHAPE
ELECTRIC FIELD STRENGTH
MEDIUM
TEMPERATURE
```
82
SEPARATES BY SURFACE CHARGE AND MOLECULAR SIZE
STRACH GEL
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SEPARATES BY MOLECULAR SIZE
CELLULOSE ACETATE
84
NEUTRAL
SEPARATES BY ELECTRIC CHARGE
NOT BIND TO PROTEIN
AGAROSE GEL
85
NEUTRAL
CHARGE AND SIZE
SEPARATES 20 PROTEIN FRACTIONS
FOR ISOENZYMES
POLYACRYLAMIDE GEL
86
COMPONENTS OF ELECTROPHORESIS
```
POWER
MEDIUM
BUFFER
SAMPLE
DETECTOR
```
87
RELATIONSHIP OF ELECTROPHORETIC MOBILITY AND NET CHARGE
DIRECTLY PROPORTIONAL
88
ELETRO MOBILITY AND ZIE AND VISCOSITY RELATIONSHIP
INVERSELY PROPORTIONAL
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IT DETERMINES THE AMOUNT OF CURRENT AND MOVEMENT OF PROTEINS FOR FIXED VOLTAGE
IONIC STRENGTH OF BUFFER
90
IONIC STRENGTH RELATION WITH CURRENT AND DISTANCE
INVERSELY PROPORTIONAL
91
RELATIONSHIP OF PH BUFFER TO MAGNITUDE OF NET CHARGE AND MOBILITY OF ELECTRIC
DIRECTLY PROPORTIONAL
92
AT PH 8.6 GAMMA GLOBULINS MOVE TOWARD THE ___
CATHODE
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IDEAL FOR SEPARATING PROTEINS OF IDENTICAL SIZES BUT WITH DIFFERENT NET CHARGE
ISOELECTRIC FOCUSING
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PRINCIPLE: PROTEINS MOVE TO ELECTRIC FIELD UNTIL PH EQUAL TO ISOELECTRIC POINT
ISOELECTRIC FOCUING
95
SUPPORT MEDIA FOR ISOELECTRIC FOCUSING
AGAROSE GEL
POLYACRYLAMIDE GEL
CELLULOSE ACETATE
96
ADVANTAGES OF ISOELECTRIC FOCUSING
RESOLVE MIXTURE OF PROCTEINS
DETECTS CK AND ALP
IDENTIFY GENETIC VARIANTS
DETECT CSF OLIGOCLONAL BANDING
97
MEASURES ABSORBANCE OF STAIN
SCAN AND QUANTITATE ELECTROPHORETIC PATTERN
DENSITOMETRY
98
MOLECULES ARE SEPARATED BY ELECTRO OSMOTIC FLOW
CAPILLARY ELECTROPHORESIS
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METHOD TO SEPARATE, DETECT,A ND IDENTIFY PROTEINS IN COMPLEX MIXTURE
WESTERN BLOTTING
100
SEPARATION OF SOLUBLE COMPONENTS BY SPECIFIC DIFFERENCES
CHROMATOGRAPHY
101
2 FORMS OF CHROMATOGRAPHY
PLANAR AND COLUMN
102
FRACTIONIZATION OF SUGAR AND AMINO ACID
PAPER CHROMATOGRAPHY
103
FOR DRUG SCREENING
COMPARING STANDARDS AND SAMPLE PLATE
THIN LAYER CHROMATOGRAPHY
104
IN DRUGS, IT IS PH DEPENDENT METHOD
EXTRACTION OF DRUGS
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IT IS A THIN PLASTIC PLATES IMPREGNATED WITH LAYER OF SILICA GEL OR ALUMINA
SORBENT
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IT IS A RELATIVE DISTANCE OF MIGRATION FROM POINT APPLICATION
RETENTION FACTOR VALUE
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FOR SEPARATION OF STEROIDS, BARBITURATES, BLOOD, ALCOHOL AND LIPIDS
BASED ON BOILING POINT
GAS CHROMATOGRAPHY
108
URINE OR BLOOD SAMPLES IN GC ARE INTROG=DUCED USING
HYPODERMIC SYRINGE OR AUTOMATED SAMPLER
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DETECTOR FOR GLC
FLAME IONIZATION
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THE RETENTION TIME OF SOLUTE IN GC OR HPLC
tR
111
MOBILE PHASE IN GAS CHROMATOGRAPHY
NITROGEN
HYDROGEN
HELIUM
ARGON
112
SEPARATION BETWEEN GASEOUS MOBILE PHASE AND LIQUID STATIONARY PHASE
GAS LIQUID CHROMATOGRAPHY
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FRACTIONIZATION AND IONIZATION
MUST BE SEPARATED BY GC FIRST
DETECT STRUCTURAL INFORMATION AND DETERMINE MOLECULAR WEIGHT
MASS SPECTROMETRY
114
- GOLD STANDARD FOR DRUG TEST
- USES ELECTRON BEAM TO SPLIT DRUG
- QUANTITATIVE
- FOR XENOBIOTICS, STEROIDS AND PESTICIDES
GC-MS
115
IT CAN DETECT 20 INBORN ERRORS FROM SINGLE BLOT
TANDEM MASS SPECTROSCOPY (MS/MS)
116
BASED ON DISTRIBUTION OF SOLUTES ON LIQUID MOBILE PHASE AND STATIONARY PHASE
LIQUID CHROMATOGRAPHY
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IS THE MOST WIDELY USED LIQUID CHROMATOGRAPHY
HPLC
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FACRTIONIZATION OF DRUGS, HORMONES, LIPIDS, CARBOHYDRATES AND PROTEINS
USES: TEMPERATURE, PRESSURE AND DETECTORS
HPLC
119
MOBILE PHASE IS MORE POLAR THAN STATIONARY PHASE
REVERSE HPLC
120
SEPARACTES ACCORDING TO SIZE AND SHAPE
LARGE MOLECULES REMAIN IN MOBILE PHASE AS TRAVELLING
GEL/ SIZE EXCLUSISON/ MOLECULAR SIEVE CHROMATOGRAPHY
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TYPE OF GEL CHROMATOGRAPHY FOR SEPARATION OF ENZYMES, ANTIBODIESM AND PROTEINS
HYDROPHILIC GEL (GEL FILTRATION)
122
TYE OF GEL CHROMATOGRAPHY SEPARATES TAG AND FATTY ACID
HYDROPHOBIC GEL (GEL PERMEATION)
123
SEPARATON OF AMINO ACIDS, PROTEINS AND NUCLEIC ACIDS
ION EXCHANGE CHROMATOGRAPHY
124
SEPARATION IS BASED ON SOLUBILITY
FOR THERAPEUTIC DRUGS AND METABOLITES
PARTITION CHROMATOGRAPHY (LIQUID-LIQUID)
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USED TO SEPARATE AND PREPARE LARGER PROTEINS AND ANTIBODIES
SEPARATE LIPOPROTEINS, CHO, AND HEMOGLOBINS
AFFINITY CHROMATOGRAOHY
126
SPEARATION BASED ON DIFFERENCES BETWEEN ADSORPTION AND DESORPTION
ADSORPTION CHROMATOGRAPHY
127
DETERMINES THE AMOUNT OF LIGHT AFTER EXCITATION BY ELECTROMAGNETIC RADIATION
FLUOROMETRY
128
MEASUERS AMOUNT OF LIGHT INTENSITY ON ZERO BACKGROUND
PHOTOMULTIPLIER TUBE
USES 2 MONOCHROMATORS
FLUOROMETRY
129
LIGHT SOURCE OF FLUOROMETRY
MERCURY ARC AND XENON LAMP
130
IT IS 1000X MORE SENSITIVE THAN SPECTROPHOTOMETER
FLUOROMETRY/ MOLECULAR LUMINISCENCE SPECTROPHOTOMETRY
131
IT IS AFFECTED BY QUENCHING
FLUOROMETRY
132
FOR MEASUREMENT OF PORPHYRINS, MAGNSEIUM, CALCIUM AND CATECHOLAMNES
FLUOROMETRY
133
IT IS THE MEASUREMENT OF CURRENT OR VOLATGE GENERATED BY ACTIVITY OF IONS
ELECTROCHEMISTRY TECHNIQUES
134
MEASURES DIFFERENCE IN VOLTAGE AT CONSTANT CURRENT
NERNST EQUATION
POTENTIOMETRY
135
REFERENCE ELECTRODE OF POTENTIOMETRY
CALOMER AND SILVER-SILVER CHLORIDE
136
EXAMPLE PH AND PCO2 TEST
POTENTIOMETRY
137
MEASURES THE ACTIVITY OF ONE ION
DEPENDS ON MEMBRANE/BARRIER COMPOSITION USED
ION SELECTIVE ELECTRODE
138
ISE ANALYZERS MEASURES
ELECTROLYTE DISSOLVED
139
IT WOULD CAUSE ERROR IN ISE MEMBRANE
PROTEIN COATING
140
MEASURES TEH AMOUNT OF ELECTRICITY AT FIXED POTENTIAL
ELECTROCHEMICAL TITRATION
DETECTED BY AMPEROMETRY
FARADAYS LAW
COULOMETRY
141
EXAMPLE: CHLORIDE TEST (SERUMA DN SEAT)
COULOMETRY
142
INTERFERENCE OF COULOMETRY
BROMIDE, CYSTEINE, AND CYANIDE
143
MEASURES TEH CURRENT FLOW BY OXIDATION REACTION
AMPEROMETRY
144
EXAMPLE: PO2, GLUCOSE, CHLORIDE AND PEROXIDASE
AMPEROMETRY
145
MEASURES DIFFERENCES IN CURRENT AT CONSTANT VOLTAGE
ILKOVIC EQUATION
POLAROGRAPHY
146
MEASURES CURRENT WHICH POTENTIAL APPLIED
PRECONCENTRATED; MINIMAL ANALYTE
VOLTAMMETRY
147
LEAD AND IRON TESTING
ANODIC STRIPPING VOLTAMETRY
