Agilent 1200 Series Evaporative Light Scattering Detector - T.E.A.M.

Agilent 1200 Series
Evaporative Light Scattering Detector
Universal detection with
optimum sensitivity for
isocratic and gradient analysis
The Agilent 1200 Series evaporative light
scattering detector (ELSD) is a powerful
tool for sensitive detection of any
compound that is less volatile than the
mobile phase. It provides an essentially
universal measurement under isocratic
and gradient conditions independent of a
compound’s absorbance, fluorescence or
electro-activity. Low temperature
evaporation technology enables detection
of semi-volatile and thermally-sensitive
compounds at low levels.
• Advanced technology for highest
sensitivity, efficiency and reproducibility
• Excellent sensitivity for low limits of
detection through low temperature
evaporation and patented gas supported
focusing within the optical head
• Applicable to semi-volatile and thermallysensitive
compounds through low
temperature technology
• Enhanced signal processing for lowest
background noise and excellent signalto-noise
ratio
• Compatible with rapid resolution liquid
chromatography (RRLC) for peak widths
of 1 second or larger
• Four nebulizers for flow rates from
5 µL/min to 5 mL/min, catering for all
important HPLC techniques
• Nebulizer and optical head designed for
optimized peak shape and peak width
• Convenient front access for inspection
and maintenance of tubing and valves
• User friendly, low maintenance system
• Operation through built-in local user
interface and Agilent ChemStation for LC
(operation through Agilent EZChrom will
be available after introduction)
How does the Agilent 1200
Series ELSD work
Step 1: Nebulize eluent and select small
droplets to minimize background noise
The nebulizer transforms the eluent from
the HPLC column into a fine vapor of
droplets. The technology in the Agilent
1200 Series ELSD provides for the selection
of droplets as a function of their size,
preventing larger droplets from entering
the evaporation or drift tube. Large droplets
cause increased background noise
because they are more difficult to
evaporate. This selection of droplets makes
detection at low evaporation temperatures
possible, resulting in low baseline noise
and excellent sensitivity, even for semivolatile
solutes.

Step 2: Evaporate at low temperature to
avoid loss of compounds
Solute molecules are obtained from the
vapor using low temperature technology in
the heated evaporation or drift tube. The
Agilent 1200 Series ELSD is designed to
evaporate high boiling mobile phases, such
as those with high aqueous concentrations,
at very low temperatures. The low
temperatures used in the standard
operating mode minimize the potential of
thermal decomposition of the compound
of interest, and makes the Agilent 1200
Series ELSD a more reliable way to detect
everything in the sample.
Step 3: Focus the solute particles, using
patented gas-supported focusing (GSF)
for enhanced signal-to-noise ratios and
less maintenance, and detect the light
scattering
Assisted by gas-supported focusing, the
solute molecules from the vapor pass
through an optical head that is designed
to measure light scattering. Gas-supported
focusing involves the addition of gas to
focus the solute particles within the optical
head for enhanced detection. The gas
surrounding the solute particles also protects
the optical head from contamination. The
magnitude of the resulting scattered light
is related to the concentration of the
compounds in the sample.
Evaporative light scattering for
detection of all peaks
In contrast to other types of detection such
as UV or fluorescence, evaporative light
scattering detectors are universal, being
capable of detecting any compound that
mV AU
1.60
1.20
1.80
0.40
0.00
4.00
3.00
1
2
Why low temperature
evaporation
The low temperature technology in the
Agilent 1200 Series ELSD minimizes
volatilization and is the key to significantly
better sensitivity.
3
4 5 6
has a lower volatility than the mobile
phase. With UV or fluorescence detection
the user can never be sure that all
compounds in the sample are detected.
This uncertainty is overcome with
evaporative light scattering detection.
2.00
7
1.00
ELSD
0.00
0.00 4.00 8.00 12.00 16.00 20.00 24.00 28.00 32.00 min
Isoflavones –
LOD = 5 ng:
1. Daidzin
2. Glycitin
3. Genistin
4. Daidzein
5. Glycitein
6. Genistein
Saponins –
LOD = 100 ng:
7. Soyasaponin I
8. Soyasaponin II
9. Soyasaponin III
10. Soyasaponin IV
0,2
0,18
0,16
0,14
0,12
0,10
0,08
0,06
0,04
8
Evaporation
at 50 0 C
Standard
9 10
UV 254 nm
Conditions:
Column: C18, 5 µm, 150 x 4.6 mm
Solvents: Water + 0.05% TFA / ACN
Flow: 1 mL/min
Pressure: 3.5 bar
Temperature: 35 °C
Analysis of biologically-active compounds in Soya with UV and evaporative light
scattering detection – the Agilent 1200 Series ELSD detects all compounds.
Evaporation
at 30 0 C
0,02
0
0 0,2 0,4 0,6 0,8 1 1,2
2
Analysis of semi-volatile caffeine with
low temperature evaporation, showing
better sensitivity than with high
temperature evaporation.

Ready for all modes of HPLC
The Agilent 1200 Series ELSD has four
nebulizers, which allows it to be used with
flow rates from 5 µL/min to 5 mL/min.
Compatibility with rapid
resolution LC
The Agilent 1200 Series ELSD is fully
compatible with the fast growing
application area of rapid resolution liquid
chromatography (RRLC) with widths of
1 second or larger. In RRLC columns
packed with sub-two micron particles are
used. The patented gas supported
focusing, which focuses the sample
molecules, and an improved optical head
design make the detector is ready for the
fast chromatograms generated in RRLC. The
column compartment temperature can be
set up to 100 °C to optimize resolution and
to minimize backpressure.
k = 1
N = 5500
k = 4
N = 12000
[mV]
7.00
6.00
5.00
4.00
3.00
2.00
Rs 1-2 = 4.5
1.00
0
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 [min]
Column: ZORBAX SB 150 mm x 500 µm, 3.5 µm
Solvents: 45 % ACN / 55 % aqueous buffer (formic acid 0.1%, pH 2.5)
Flow:
45 µL/min
Column temperature: 90 °C
Elution order: Nitrazepam, flunitrazepam, diazepam, ibuprofen,
mefenamic acid (40–400 ng injected)
Nebulizer:
Micro-flow
Analysis of pharmaceutical drugs with microliter flow and micro-flow nebulizer.
The chromatogram shows the baseline separation of four pharmaceutical drugs on a
500 µm i.d. column and flow rate of 45 µL/min. Using the micro-flow nebulizer results
in excellent peak shapes and plate numbers.
[mV]
700
600
500
400
300
200
100
0
0.133
0.173
0.242
0.357
0 0.1 0.2 0.3 0.4 0.5
[min]
Column: ZORBAX SB-C18, 2.1 x 50 mm, 1.8 µm
Solvents: Water +0.1 % TFA/ACN +0.1 % TFA
Flowrate: 1 mL/min
Column temperature: 40 °C
Detector pressure: 5 bar
Evaporation temperature: 26 °C
RRLC of semi-volatile parabens at 40 °C
column temperature with the Agilent 1200
Series ELSD.
Volts
0.018
0.016
0.014
0.012
0.010
0.008
0.006
0.004
0.002
0 2 4 6
Column: C18, 150 x 1 mm, 5 µm
Solvents: Water/Methanol (25/75)
Flow:
50 µL/min
Detector pressure: 3.5 bar
Evaporation
temperature: 28 °C
Sample:
Methylparaben, ethylparaben,
propylparaben, butylparaben
(8 ng each)
Paraben S/N
Methyl- 4
Ethyl- 6
Propyl- 17
Butyl- 27
Analysis of semi-volatile parabens with an evaporation temperature of 28 °C.
3

Optimized for high performance
detection
The Agilent 1200 Series ELSD provides
unmatched sensitivity for semi-volatile and
non-volatile compounds. An example for
the excellent sensitivity is the analysis of
the semi-volatile parabens. Just 8 ng of
each paraben were injected, resulting in
signal-to-noise ratios between 4 and 27.
The Agilent 1200 Series ELSD can achieve
limits of detection (LOD*) of 2 ng for
glucose (a typical non-volatile analyte), and
1 ng for caffeine (a typical semi-volatile
analyte).
Control and data evaluation
The Agilent 1200 ELSD can be controlled
through the built-in local keyboard. For
more comfortable operation, control of the
detector is fully integrated in the Agilent
ChemStation. Control through Agilent
EZChrom Elite is expected to be available
early 2008.
*Reference conditions
Column: ZORBAX Eclipse Plus C18,
2.1 x 30 mm, 3.5 µm
Solvents:
100 % water (glucose analysis),
water/acetonitrile 70/30
(caffeine analysis)
Flow:
0.2 mL/min
Column temp.: 22 °C
Evaporation temp.: 50 °C
(glucose analysis),
30 °C (caffeine analysis)
Nebulizer: Semi-micro flow (option 016)
**Reference conditions
Eluent flow: 0 ml/min; Standard-flow evaporator
temperature: 50 °C; Nebulizer gas: Nitrogen flow at
3.5 bar; Gain: 12; Noise determination: Mean value of
10 segments of 1 minute each, for each segment
measure the difference between the lowest and
highest peak (peak-to-peak) and divide by 2.
Agilent ChemStation control menu showing ELSD signal and instrument set points.
Specifications
Detection:
High sensitivity photomultiplier
Light Source:
High efficiency Blue LED (470nm)
Temperature range (evaporator) Ambient to 100 °C
Flow rate:
5 µL/min – 5 mL/min
Nebulizers:
Large flow (optional): 1–5 mL/min (useable flow range)
Standard flow (included): 0.2–2.5 mL/min (usable flow range)
Semi-micro flow (optional): 0.02–1.2 mL/min (usable flow range)
Micro flow (optional):
0.005–0.08 mL/min (usable flow range)
Short-term noise:
< 2 mV**
Drift:
< 1 mV/h**
Signal Output:
0-1 V (analog), RS 232 (digital)
Gain Settings: 2 11 (2048)
Communications:
RS-232, Contact closure for ready, auto-zero, APG remote
Control and data evaluation: Built-in local controller
Agilent ChemStation revision B.03.02 or higher
Agilent EZChrom Elite 3.3 (available early 2008)
Context-sensitive help: Agilent ChemStation
Gas flow control:
Manual and computer-controlled (power down) nebulization gas
flow and patented auxiliary gas flow
Gas consumption:
Micro-flow, semi-micro and standard flow nebulizers less than 3 L/min;
Large flow nebulizer 4 L/min
Environment:
5-40 °C at < 95 % humidity
Dimensions: Width: 250 mm (10 in)
Height: 450 mm (18 in)
Depth: 550 mm (22 in)
Weight: 18.5 kg (40 lb)
Ordering information
www.agilent.com/chem/1200
© Copyright 2007 Agilent Technologies
Published December 1, 2007
Publication Number 5989-7473EN
Description
Order No.
Agilent 1200 Series evaporative light scattering detector,
including standard nebulizer (usable flow rate 0.2-2.5 mL/min),
RS-232 and APG remote start cables, user’s guide.
G4218A
Large flow nebulizer (1-5 mL/min) G4218A #020
Semi-micro flow nebulizer (0.02-1.2 mL/min) G4218A #016
Micro-flow nebulizer (0.002-0.08 mL/min) G4218A #010
Gas regulator with 0.01 µm filter G4218A #030