HPLC DETECTORS

Characteristics
Low drift of base line and low background
noise (LOD)
High sensitivity
Rapid response
Large
linear dynamic range (> 105)
Low dead volume
 Types of detectors
1. BULK PROPERTIES DETECTORS
measure a characteristic of the MP influenced by the presence of
solute. Not selective
The signal is proportional to the flow of the MP (m/t)
Universal, less sensitive, small linear dynamic range.
No gradient elution
1. Refraction index
2. Electrochemical
3. Evaporative light scattering

2. SOLUTE PROPERTIES DETECTORS

Measure properties which are characteristic of the solute (not all
solute are detected). Selective.
High sensitivity.
The signal is proportional to the solute concentration

1. UV absorption detector
2. Fluorescence
HPLC Bulk Property Detectors

Refractive Index Detector

Fresnel refractometer

1. Fuente de radiacin
visible
2. Doble abertura
3. Lente
4. Prisma
5. Lente (interfaz entre
prisma- liquido)
 HPLC Bulk Property Detectors

Refractive Index Detector

Plus:
1) Measures a bulk property
2) Nearly Universal (different RI than mobile phase)
3) Comparable response for different analytes
4) Detects species with no chromophores

Minus:
1) Temperature dependent
2) Poor sensitivity (LOD 100 ng)
3) No gradient elution
 HPLC Bulk Property Detectors
Evaporative Light Scattering Detector

Solid state laser

 HPLC Bulk Property Detectors
Evaporative Light Scattering Detector

Plus:
1) Measures a bulk property
2) Nearly Universal (must be non-volatile)
3) Does not detect liquids (gradients are ok)
4) Detects species with no chromophores

Minus:
1) Signal not linear with concentration
2) Fair sensitivity (LOD 1 ng)
3) No salts or buffers in mobile phase
HPLC Bulk Property Detectors
Conductivity Detector
 HPLC Bulk Property Detectors
Conductivity Detector

Plus:
1) Measures a bulk property
2) Nearly Universal (must be ionic)
3) Common for Ion Exchange LC
4) Detects species with no chromophores
5) Simple, robust

Minus:
1) Fair sensitivity (LOD 1 ng)
2) No salts or buffers in mobile phase
3) Gradients a challenge
HPLC Specific Property Detectors

Conventional UV-Vis Absorption Detector

Photodiode 1

Photodiode 2
Hg Pen
Lamp
254 nm
 HPLC Specific Property Detectors

Multi-wavelength UV-Vis Absorption Detector

Deuterium Photodiode
Lamp Array
 HPLC Specific Property Detectors

Multi-wavelength UV-Vis Absorption Detector

 HPLC Specific Property Detectors

Multi-wavelength UV-Vis Absorption Detector

Plus:
1) Measures a specific property
2) Nearly Universal (must have chromophore)
3) Most common of all detectors (~75%)
4) Potential to provide qualitative info.
5) Simple, robust

Minus:
1) Fair sensitivity (LOD 1 ng)
2) Expensive with PDA, limited with Hg Lamp
3) Misses some important analytes
HPLC Specific Property Detectors

Fluorescence Detector
 HPLC Specific Property Detectors

Fluorescence Detector
 HPLC Specific Property Detectors

Fluorescence Detector

Plus:
1) Measures a specific property
2) Highly Selective (must fluoresce)
3) Second most common of all detectors (~15%)
4) High sensitivity (LOD 0.01 ng)
5) Can interrogate very small volumes

Minus:
1) Not Universal
2) Limited Applications
HPLC Specific Property Detectors

Electrochemical Detector
 HPLC Specific Property Detectors

Electrochemical Detector

Plus:
1) Measures a specific property
2) Highly Selective (depends on reduction potential)
3) High sensitivity (LOD 0.01 ng)

Minus:
1) Not Universal
2) Must have electrolyte in mobile phase
3) Mobile phase must be aqueous
4) Gradients not possible
HPLC Element Specific Detectors

ICP Detector Detector

 HPLC Element Specific Detectors

ICP Detector Detector

Plus:
1) Can be Highly Selective
2) Can be nearly Universal
3) Can have single sensitivity for all analytes

Minus:
1) $$$
2) Mobile phase must be aqueous
4) Mobile phase must not contain element of interest
UV-Vis detection of sugars in juice
UV-Vis detection of sugars in juice
ICP Carbon detection of sugars in juice
ICP Carbon
detection of sugars
in orange juice
ICP Carbon detection of amino acids
ICP Carbon detection of amino acids
HPLC Derivatization Detectors

Tag analytes with fluorophore

Before or After Separation
HPLC Derivatization Detectors

Tag analytes with fluorophore

Before or After Separation
 HPLC Indirect Detectors
Detect the change in a constant signal
as the analyte passes
Recent HPLC Detector
Hollow Cathode Detector