Agilent 1290 Infinity Evaporative Light Scattering Detector (ELSD) For Low Molecular Weight Compounds Introduction Evaporative light scattering detectors (ELSDs) are ideal for detecting analytes with no UV chromophore as they do not rely on the optical properties of a compound. The Agilent 1290 Infinity Evaporative Light Scattering Detector (ELSD) is a high performance ELSD, benefitting from almost 20 years experience in the design and manufacture of ELSDs The 1290 Infinity ELSD is the only ELSD that delivers subambient operation, for unrivalled detection of thermally labile analytes. Designed for the analysis of all compounds, the 1290 Infinity ELSD delivers evaporation down to 10 °C, providing maximum sensitivity for compounds with significant volatility below 30 °C. Programmable control of gas flow and temperature during an injection eliminates solvent gradient effects and maximizes response for improved accuracy. To detect everything you inject with high sensitivity, the 1290 Infinity ELSD is the ELSD of choice for all applications. Key Benefits • High sensitivity provides superb responses for all compounds, down to low nanogram levels. • Subambient operation using a Peltier cooled evaporation tube delivers temperatures down to 10 °C, preventing degradation of the thermally labile compounds that other ELSDs cannot detect. • Real-time control during an injection using programmable Dimension Software maintains maximum sensitivity throughout the run. • Real-time gas programming using Dimension Software eliminates solvent enhancement effects during gradient elution, for excellent quantification. • Low dispersion and high-speed data output rates are the perfect match for fast LC applications. • Superb reproducibility below 2% gives reliable and accurate results. Data Sheet
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IntroductionEvaporative light scattering detectors (ELSDs) are ideal for detecting analytes withno UV chromophore as they do not rely on the optical properties of a compound.The Agilent 1290 Infinity Evaporative Light Scattering Detector (ELSD) is a highperformance ELSD, benefitting from almost 20 years experience in the design andmanufacture of ELSDs The 1290 Infinity ELSD is the only ELSD that deliverssubambient operation, for unrivalled detection of thermally labile analytes. Designedfor the analysis of all compounds, the 1290 Infinity ELSD delivers evaporation downto 10 °C, providing maximum sensitivity for compounds with significant volatilitybelow 30 °C. Programmable control of gas flow and temperature during an injectioneliminates solvent gradient effects and maximizes response for improved accuracy.To detect everything you inject with high sensitivity, the 1290 Infinity ELSD is theELSD of choice for all applications.
Key Benefits• High sensitivity provides superb responses for all compounds, down to low
nanogram levels.
• Subambient operation using a Peltier cooled evaporation tube deliverstemperatures down to 10 °C, preventing degradation of the thermally labilecompounds that other ELSDs cannot detect.
• Real-time control during an injection using programmable Dimension Softwaremaintains maximum sensitivity throughout the run.
• Real-time gas programming using Dimension Software eliminates solventenhancement effects during gradient elution, for excellent quantification.
• Low dispersion and high-speed data output rates are the perfect match for fastLC applications.
• Superb reproducibility below 2% gives reliable and accurate results.
Data Sheet
effects to give a constant responseacross a gradient. Control and digitaldata collection come as standard formultivendor platforms so there is noneed for an analog to digital converter.On-the-fly adjustment of light sourceintensity can save time during a run. Allthis in the smallest footprint available.Being complementary to LC/MS, andoffering unrivalled flexibility andsensitivity, the 1290 Infinity ELSD is theELSD of choice for the most demandingapplications.
The Agilent Family of EvaporativeLight Scattering DetectorsEvaporative light scattering detection isthe powerful alternative to RI and UV,inject it and detect it, right down to lownanogram levels. ELSDs provide auniversal response independent of theoptical properties of the analyte sothere is no loss of importantinformation.
Agilent ELSDs can be used for a widerange of analytical techniques,including LC (analytical andpreparative), LC/MS, SFC, highthroughput screening, GPC/SEC, TREF,and GPEC. The range of applicationareas is equally broad, encompassingpharmaceuticals, nutraceuticals,combinatorial libraries, carbohydrates,lipids, phopspholipids, triglycerides,fatty acids, amino acids, polymers, andsurfactants.
Since the 1290 Infinity ELSD deliverssubambient evaporation down to 10 °C,it provides maximum sensitivity forcompounds with significant volatilitybelow 30 °C. For nonvolatile compounds,where temperatures of 100 °C or aboveare required, the 1260 Infinity ELSD isthe instrument of choice.
Key Benefits (Continued)• Multivendor software control and
data acquisition using AgilentChemStation chromatography datasystem, and other vendors’interfaces, eliminates the need foran analog to digital converter.
• Rapid heating and cooling of theevaporator tube minimizesequilibration time.
• Full DMSO transparency ensuresthat responses from early elutingcompounds are not hidden.
• Fully integrated with all Agilentanalytical and preparative LCsystems for the completechromatographic solution.
• Complementary to LC/MS.
System DetailsThe 1290 Infinity ELSD deliverssubambient evaporation down to 10 °C,providing maximum sensitivity forcompounds with significant volatilitybelow ambient temperature. Theinstrument benefits from fast dataoutput rates and extremely lowdispersion for fast LC, and delivers auniversal response down to the low-nanogram range for truly representativeanalysis. Reproducibility is less than 2%for improved consistency of results. The1290 Infinity ELSD offers real time gasmanagement that eliminates solvent
All Agilent ELSDs are rigorously testedto a high specification, and detailedIQ/OQ documentation is included withevery unit before they are shipped. Inaddition, Agilent detectors can beincorporated into any LC system, acomplete Qualification Workbook isprovided to help you.
Uniform Mass ResponseAgilent ELSDs are not dependent on acompound’s optical properties so theELSD provides a more uniformresponse than UV-VIS, making it theideal detector for purity analysis orwhere calibration standards are notavailable.
1. Nebulization–Using an inert gasstream to form a plume of uniformlysized droplets.
2. Evaporation of the eluent–Generatinga plume of nonvolatile solute particles.
3. Optical detection–Where theintensity of scattered light isproportional to the mass of solutepassing through the optical chamber.
NebulizationEfficient nebulization using low gasflow rates is a feature of Agilent ELSDs.Independent nebulizer temperaturecontrol and digital gas flow controlprovide excellent stability andreproducibility. Baseline noise isminimized by the removal of any poorlynebulized eluent through a drain port.
EvaporationThe nebulized stream passes through anindependently temperature-controlledevaporator tube where solvent isremoved at high temperature, leavingthe less volatile solute particles behind.
Gas inlet
Liquid wasteEluentinlet Light source
Detector
The 1290 Infinity ELSD featurespatented1 gas flow control technologywith a short evaporator tube that givesan extremely low swept volume forminimal peak dispersion. This providesmaximum resolution from theseparation, especially important forwork with small columns.
1 UK Patent 0304253.8, US Patent 6/0238744
Optical detectionThe solute particles are detected asthey pass through the optical chamber.The high power LED and advanceddesign of the electronics delivers maximum sensitivity.
Unrivalled Uniform ResponseAcross a GradientThe 1290 Infinity ELSD is unique in itsability to control gas flows during a runto produce a uniform detector responseacross a gradient. This real-time benefitoffers an alternative approach to mobilephase compensation when quantifyingunknowns under gradient conditions.
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Sample: ParabensColumn: C18 2.1 × 150 mm, 5 µmEluent A: WaterEluent B: AcetonitrileGradient: 5% to 70% B in 5 minutes;
70% to 95% B in 2 minutesFlow rate: 0.2 mL/minInjection volume: 10 µL Detector: Agilent 1290-ELSD (neb = 30 °C,
evap as shown, gas = 1.4 SLM)
min1513.812.611.410.29
20 °C
30 °C
40 °C
1 23 4
5
1 23
45
34
5
Figure 4Improved detection of parabens using subambient ELSD with the Agilent 1290 InfinityELSD (parabens mixture injected at 100 ngexcept heptyl at 160 ng).
Column: C18 2.1 × 150 mm, 5 µmEluent A: WaterEluent B: AcetonitrileGradient: 5% to 95% B in 10 minutesFlow rate: 1.0 mL/minInjection volume: 10 µL (every minute)Detector: Agilent 1290-ELSD (neb = 30 °C,
evap = 30 °C)
min0 2 4 6 8 10
Gas control on
Gas control off
Figure 3Real-time gas control using DimensionSoftware provides unrivalled uniformity acrossa gradient.
Fast LC
The excellent baseline stability acrosssteep gradients and low dispersioncharacteristics combined with fast dataoutput rates makes the 1290 InfinityELSD ideal for fast LC.
Sample: Tertiary aminolsColumn: C18 4 × 50 mm, 5 µmEluent A: Water + 0.1% Formic acidEluent B: Acetonitrile + 0.1% Formic acidGradient: 5% to 95% B in 5 minutesFlow rate: 2.5 mL/minInjection volume: 10 µL Detector: Agilent 1290-ELSD (neb = 40 °C,
evap = 50 °C, gas = 1.5 SLM)
Figure 2The Agilent 1290 Infinity ELSD provides a stablebaseline even across steep, high speedgradients.
0 min 3
Subambient ELSDThe 1290 Infinity ELSD’s unique cooledevaporation zone provides subambientoperation to deliver unrivalled detectionof low molecular weight compoundsnot seen by other ELSDs.
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Superb RSDExcellent reproducibility below 2% givesreliable and accurate results. You canhave complete confidence in your data.
min543210
50
45
40
Tem
p °C
35
30
Figure 6Fast temperature equilibration with the Agilent 1290 Infinity ELSD.
Figure 5Fifty caffeine injections produce an RSD of 1.27%, demonstrating the superb reproducibilityof the Agilent 1290 Infinity ELSD.
Sample: 2 mg/mL in DMSOColumn: 4.6 × 150 mm, 5 µm Eluent A: 0.1% HFBA in WaterEluent B: 0.1% HFBA in AcetonitrileGradient: 5% to 30% B in 10 minutes,
30% to 80% B in 5 minutesFlow rate: 1.0 mL/minInjection volume: 20 µLDetector: Agilent 1290 Infinity ELSD
(neb = 25 °C, evap = 25 °C, gas as shown)
DMSO
Gas = 1.2 SLM
Gas = 1.8 SLM
0 min 15
Figure 7Increasing gas flow on the Agilent 1290 InfinityELSD gives DMSO transparency.
Fast Temperature SwitchingFor higher throughput of samples, theunique 1290 Infinity ELSD features rapidtemperature equilibration. The detectorcools from 50 °C to 30 °C, andvice versa, in just over 5 minutes. Inaddition to the enhanced detection ofsemivolatile compounds, thetemperature controlled evaporator tubeon the 1290 Infinity ELSD allows theuser to rapidly change betweentemperature programs.
DMSO Transparency for HighThroughput ScreeningThe vast majority of combinatoriallibraries screened by high throughputmethods are stored in dimethylsulfoxide(DMSO). UV and mass spectrometrywill miss potential drug compoundsthat have no UV chromophore, so ELSDis commonly used. However, at the lowevaporator temperatures required todetect volatile components, the presence of DMSO in the sample canmask the response of early eluting compounds. In the 1290 Infinity ELSD,the addition of a carefully controlledstream of gas to the evaporation stepenables complete removal of the DMSOto take place without increasing theoperating temperature.
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Use of the sophisticated 1290 InfinityELSD software ensures improveddetection of compounds not normallyamenable to LC/MS.
Choosing Detector SettingsThe unique patented gas flow operationof the 1290 Infinity ELSD makes itmobile phase independent. Instead,
Adding Value to LC/MSThe 1290 Infinity ELSD is fullycomplementary to LC/MS. Theinstrument can provide better data andsupport information for compounds thatdo not perform well on massspectrometers, such as haloperidol,enhancing the value of your LC/MSsystem.
min107.552.5
1
2
3Peak identification1. Haloperidol2. Haloperidol metabolite3. Haloperidol metabolite(1 µg of each on column)
LC/MS
Figure 8Coupling the Agilent 1290 Infinity ELSD withLC/MS reveals much greater detail in theanalysis of haloperidol and its metabolites.
min107.552.5
1
2
3
Agilent 1290 Infinity ELSD
settings are optimized according to thethermal sensitivity of compoundsinjected.
As a general rule, all solvents can beevaporated with nebulizer andevaporator temperatures at 30 °C and agas flow of 1.6 SLM. This can be usedas a starting point and adjustmentsmade according to the sample injected(if necessary), see Table 1.