Tips & Tricks Dwn_ds

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www.polymerlabs.com/elsd DETECT EVERYTHING YOU INJECT WITH HIGH SENSITIVITY Evaporative Light Scattering Detectors FOR HPLC LC-MS SFC HIGH THROUGHPUT SCREENING GPC TREF GPEC Now a part of Varian, Inc. PL-ELS 2100 PL-ELS 1000 PL-ELS 2100 Ice NEW

Transcript of Tips & Tricks Dwn_ds

Page 1: Tips & Tricks Dwn_ds

www.polymerlabs.com/elsd

D E T E C T E V E R Y T H I N G Y O U I N J E C T W I T H H I G H S E N S I T I V I T Y

Evaporative Light Scattering Detectors

F O R H P L C ■ L C - M S ■ S F C ■ H I G H T H R O U G H P U T S C R E E N I N G

G P C ■ T R E F ■ G P E C

Now a part of Varian, Inc.

PL-ELS 2100

PL-ELS 1000

PL-ELS 2100 Ice

NEW

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It is well recognized that Evaporative Light ScatteringDetection (ELSD) can outperform traditional detectors whenanalyzing non-chromophoric samples by HPLC as thedetection method does not rely on the optical properties ofthe analyte. Furthermore, an ELSD detects all compoundsless volatile than the mobile phase and, with advanceddesign features for low temperature operation, the benefitsof ELSD compared to UV or refractive index detection nowapply to an even wider range of HPLC applications.

Evaporative Light Scattering Detectors from Polymer LaboratoriesPolymer Laboratories (PL), now a part of Varian, Inc., is a worldleader in the design, build, service and support of evaporativelight scattering detectors with over 17 years’ experience in thisfield. PL’s instruments have evolved, and continue to develop,through listening to the requirements of our customers.

Polymer Laboratories presents three models of ELSD, eachoffering high sensitivity for a wide range of analytes and analysisconditions, and each designed to address your particularapplication area. Choose from the PL-ELS 2100 for regular HPLC,the new sub-ambient PL-ELS 2100 Ice (Integrated CooledEvaporator) for the detection of very low boiling point compoundsor the PL-ELS 1000 for routine HPLC and GPC. Whichever optionyou choose, PL will deliver you high quality, high sensitivity, stateof the art evaporative light scattering detection.

PL-ELS 2100 PL-ELS 2100 Ice PL-ELS 1000

Designed and built to the highest standards, Polymer Laboratories’detectors offer considerable benefits compared to other ELSDson the market:

■ Extremely small footprint to maximize available bench space

■ Wide gradient compatibility, no limitation on choice of solvents

■ High sensitivity, extremely low limits of detection (LoD)

■ Wide application range to cover very low to high molecular weight compounds

■ Low dispersion, minimal peak broadening for maximum resolution

■ Rapid equilibration, fast set-up and high productivity

■ Advanced, easy to use instrument control with built-in safety

ELSD detectors from PL offer the widest flexibility and deliveroptimum performance for all applications.

Detect Everything You Inject with High Sensitivity

PL-ELS 2100 PL-ELS 1000 PL-ELS 1000µ

HPLC analysis of non-volatile compounds ✔ ✔ ✔

HPLC analysis of semi-volatile compounds ✔ ✔ ✘

Ambient GPC analysis ✔ ✔ ✔

High temperature GPC analysis ✘ ✘ ✔

PL-ELS 2100 PL-ELS 2100 PL-ELS 1000Ice

Application Areas

■ Pharmaceuticals

■ Nutraceuticals

■ Combinatorial libraries

■ Carbohydrates

■ Lipids

■ Phospholipids

■ Triglycerides

■ Fatty acids

■ Amino acids

■ Polymers

■ Surfactants

Analytical Techniques

■ HPLC - analytical, microbore, preparative

■ LC-MS

■ High throughput screening

■ GPC/SEC

■ High temperature GPC

■ TREF

■ GPEC

Universal ResponseELSD does not rely on the optical properties of the analyte makingit an ideal detector for many compounds, including those with noUV chromophore or widely differing extinction coefficients. WhenUV detection is used to monitor the separation of equal amountsof aspirin and phenacetin, different responses are recorded dueto variable extinction coefficients at 280nm. Clearly, a more uniformresponse is obtained when the PL-ELS detectors are used.

Sample: 1. Aspirin, 2. PhenacetinColumn: PLRP-S 100Å 5µm, 150x4.6mmEluent: 0.1% TFA in 50% Water : 50% ACNFlow Rate: 0.5ml/minDetectors: PL-ELS 2100 (neb=30°C, evap=30°C, gas=1.6 SLM)

UV, 280nm

KEY1. Aspirin2. Phenacetin

100 min

PL-ELS 2100

UV, 280nm

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2

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ELSD - The Ideal Substitute for, or Supplement to, YourExisting HPLC DetectorsEvaporative light scattering detection has become a regulardetection method for liquid chromatography, offering key benefitsover more conventional UV and RI detectors:

■ Does not require the sample to have a chromophore

■ Universal response, no effect of extinction coefficient or dn/dc

■ Fully gradient compatible, no limitation on solvent choice

■ Better alternative to UV detection at low wavelengths

■ No need for pre- or post-column derivatization

■ Excellent baseline stability

Better Alternative to UV Detection at Low WavelengthsEvaporative light scattering detection is a more favorablealternative than UV detection for the analysis of compounds thatrequire low wavelength analysis (e.g. 230nm) or possess a weakchromophore. These compounds have a low sensitivity to UVdetection and suffer from baseline instability under gradientelution. The benefit of ELSD for these types of compounds ishighlighted in Figure 1, which compares UV and ELS detectionfor the analysis of a black cohosh sample.

Compounds: 1. Cimiracemoside, 2. Actein, 3. 27-deoxyacteinSample: Black Cohosh TabletColumn: Inertsil C18 5µm, 150x4.6mmEluent A: 0.1% Formic acid in WaterEluent B: ACNGradient: 30-40% B in 30 mins, 40-60% B in 30 mins Flow Rate: 1.0ml/minInj Vol: 20µlDetectors: PL-ELS 2100 (neb=30°C, evap=50°C, gas=1.4 SLM)

UV, 230nm

KEY1. Cimiracemoside2. Actein3. 27-deoxyactein

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600 min

PL-ELS 2100

UV, 230nm

A Valuable Addition to Your LC-MS SystemAs the operational requirements are very similar, methodsdeveloped on PL’s evaporative light scattering detectors can bedirectly transferred to MS, saving valuable LC-MS methoddevelopment time. ELSDs from Polymer Laboratories can be usedin parallel with any standard LC-MS system to obtain concentrationinformation for all compounds, whether they are UV active or not.

Samples: 1. Verapamil 2. DiazepamColumn: Commercially available C8 3µm, 150x2.1mmEluent A: 0.1% Formic acid in WaterEluent B: 0.1% Formic acid in ACNGradient: 20-80% B in 5 minsFlow Rate: 0.3ml/minInj Vol: 10µlDetectors: MS, UV, PL-ELS 2100 (neb=50°C, evap=50°C, gas=1.6 SLM)KEY1. Verapamil2. Diazepam

■ LC-MS traceshows 1:3 ratio

■ UV traceshows 1:10 ratio

■ ELS shows thetrue ratio of 1:1

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PL-ELS 2100

UV

MS150 min

DMSO

DMSO Transparency for High Throughput ScreeningThe vast majority of combinatorial libraries screened by highthroughput methods are stored in Dimethylsulfoxide (DMSO). UV and Mass Spectrometry will miss potential drug compoundsthat have no UV chromophore, so ELSD is commonly used.However, at the low evaporator temperatures required to detectvolatile components, the presence of DMSO in the sample canmask the response of early eluting compounds. In the PL-ELS2100, the addition of a carefully controlled stream of gas to theevaporation step enables complete removal of the DMSO to takeplace without increasing the operating temperature. The benefitsof this DMSO transparency are highlighted in the figure below.

■ The DMSO response masks the fast eluting peak

■ Increasing the gas flow to 1.8 SLM gives DMSO transparency

Sample: 2mg/ml in DMSOColumn: Inertsil 7 ODS-3 5µm,

150x4.6mmEluent A: 0.1% HFBA in WaterEluent B: 0.1% HFBA in ACNGradient: 5-30% B in 10 mins,

30-80% B in 5 mins Flow Rate: 1.0ml/minInj Vol: 20µlDetector: PL-ELS 2100

(neb=25°C, evap=25°C, gas as shown)

gas=1.8 SLM

gas=1.2 SLM

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ELSDs from Polymer Laboratories - Better By Design

Evaporative light scattering detection involves a three stageprocess:

NebulizationEfficient nebulization using low gas flow rates is a feature of PL’sdetectors. Independent nebulizer temperature control and digitalgas flow control provide excellent stability and reproducibility. Any poorly nebulized eluent is removed via a drain port tominimize baseline noise.

EvaporationThe nebulized stream passes through an independentlytemperature controlled evaporator tube where the solvent isremoved, leaving the less volatile solute particles. PL’s detectorsfeature the shortest evaporator tubes resulting in extremely lowswept volume and minimum peak dispersion. The result is thatevaporative light scattering detectors from Polymer Laboratoriesdeliver you the maximum resolution from your separation,especially important for work with small column volumes.

gas inlet

liquid waste

light source

detector

eluent inlet

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Optical DetectionThe solute particles are detected as they pass through the opticalchamber. Advanced design of the optical and electroniccomponents ensures that PL’s detectors deliver maximumsensitivity with minimum baseline noise.

Sample: Fentanyl Column: Commercially available C18 5µm, 50x2.1mmEluent A: 60% 0.25M Ammonium acetate in Water, 40% ACNFlow Rate: 0.25ml/minInj Vol: 10µlDetector: PL-ELS 2100 (neb=40°C, evap=70°C, gas=1.2 SLM)

■ Achieve low ng levels of detection on column

40 min

100ng on column

20ng on column

Easy and Safe to UsePL’s detectors offer both onboard operation via a keypad anddisplay or advanced instrument control using a Windows PCbased graphical user interface. Up to 10 pre-set methods can bestored, and the control software provides the facility forautomation and logging procedures. In addition, the controlsoftware can be used to run a series of diagnostic checks to testfor system compliance and validate performance, as well as toprovide interaction with the multiple safety features built into thedetectors.

Instrument Qualification - IQ, OQAll Polymer Laboratories’ detectors are rigorously tested to a highspecification before they are shipped, and detailed IQ, OQdocumentation is included with each unit. In addition, PL’sdetectors can be incorporated into any HPLC system, and acomplete Qualification Workbook is provided to assist you.

Galaxie Control and Data Handling1

The PL-ELS 2100 can be controlled via the Galaxie Workstationfrom Varian, Inc. to provide integrated control of your system,data analysis and reporting. The Galaxie Workstation alsocontains a suite of features that allows customers to comply withFDA 21 CFR Part 11 regulations.

1Available from Summer 2006

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Analysis of HerbicidesUnlike the majority of pesticides, the application of gaschromatography (GC) to the analysis of phenylurea pesticides isdifficult because they are thermally unstable and degrade rapidlyto isocyanates and amines. An alternative approach for thermallysensitive compounds is to use HPLC coupled with lowtemperature evaporative light scattering detection. The PL-ELS2100 from Polymer Laboratories is designed to evaporate highlyaqueous mobile phases at ambient temperature in order tomaximize the response of semi-volatile compounds. The benefitsof this design are highlighted in this analysis of 10 phenylureaherbicides.

Sample: Phenylurea-herbicide mixtureColumn: Commercially available C18 5µm,

250x4.6mmEluent A: WaterEluent B: ACN Gradient: 10-80% B in 40 minsFlow Rate: 0.7ml/minInj Vol: 20µlDetector: PL-ELS 2100 (gas=1.6 SLM)

■ By operating the PL-ELS 2100 at ambient temperature, sample

Pharmaceutical ExcipientsCyclodextrins are used to facilitate the dissolution of hydrophobicdrugs; the manner in which they interact is important in developinga successful pharmaceutical formulation. UV detection is thestandard method for pharmaceutical analysis, however,cyclodextrins do not possess a UV chromophore. The PL-ELS 2100provides a beneficial alternative, because it is not dependent on acompound’s optical properties. In this example, the PL-ELS 2100is able to detect the excipient cyclodextrins along with the targetdrug ibuprofen

Column: Commercially available C18 5µm, 150x4.6mmEluent A: WaterEluent B: ACNGradient: 50-95% B in 5 minsFlow Rate: 1.0ml/minInj Vol: 20µlDetectors: PL-ELS 2100 (neb=30°C,

evap=50°C, gas=1.0 SLM)UV, 221nm

KEY1. α-cyclodextrin2. ß-cyclodextrin3. Ibuprofen

■ The PL-ELS 2100 can provide the true composition of pharmaceutical formulations in a single chromatogram

PL-ELS 2100 - Surpasses other ELSDs for High Sensitivity HPLC AnalysesThe PL-ELS 2100 represents the leading edge of ELSDdesign, providing optimum performance across adiverse range of HPLC applications. Its patented designpermits operation at ambient temperature, to facilitatethe detection of semi-volatile compounds not seen by other ELSDs.

In evaporative light scattering detection, the eluent is removed asit passes through the evaporator tube. The temperature of thistube is critical in optimizing the detection of small molecules. If the temperature is too high, thermally labile compounds in thesample may evaporate along with the eluent, and will not bedetected. The PL-ELS 2100 can evaporate high boiling solvents atvery low temperatures, for example a mobile phase consisting of100% water at 30°C. The novel design of the PL-ELS 2100provides a level of performance for semi-volatile compounds thatis superior to that which can be obtained from competitors’ELSDs.

90 min

1

2 3 4

35oC

Ambient

50oC

400 min

400 min 400 min

Analysis of Pharmaceutical CompoundsIn this analysis, peak 2, indapamide, is essentially non-volatile butthe remaining three peaks (acetanilide, ibuprofen anddibutylphthalate) are semi-volatile to different degrees. The effectof operating temperature on the recovery of these three semi-volatile compounds is clearly illustrated. With the nebulizerand evaporator running at the same temperature, 50°C, peak 2gives a large response but the other more volatile componentsgive very poor peak response due to loss through evaporation.As the nebulizer/evaporator temperatures are reduced to 35°Cand further, when they are switched off and the instrument isrunning at ambient temperature (nominally 26°C), the detectorshows much better recovery of the semi-volatile components withno deterioration in baseline stability.

Column: Commercially available C18 5µm, 150x4.6mm Eluent A: Water + 0.1% TFAEluent B: ACN + 0.1% TFA Gradient: 60-90% B in 5 minsFlow Rate: 1.0ml/minInj Vol: 20µlSample Conc: 1mg/mlDetector: PL-ELS 2100

KEY1. Acetanilide2. Indapamide 3. Ibuprofen4. Dibutylphthalate

50°C

25°C

70°C

21

3

120 min

PL-ELS 2100

UV, 221nm

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Triglycerides in Plant Oil using Evaporative Light Scattering DetectionPlant oils contain complex mixtures of triglycerides which need tobe analyzed using gradient elution. Refractive index detection isnot gradient compatible, whereas UV lacks sensitivity due to thelow wavelengths required. Evaporative light scattering detection isuniversal, and is not dependent on the optical properties of thecompound, making it a better alternative to UV detection.

Optimum resolution and reduced run time is easily achieved withthe PL-ELS 2100 due to its compatibility with a wide range ofsolvents and insensitivity to solvent gradients.

Supercritical Fluid Chromatography (SFC) of Polyethylene GlycolsPolyethylene glycols (PEGs) are low molecular weight polymersthat are widely used as excipients or drug delivery agents in thepharmaceutical industry, as well as additives in cosmetics andhome care products.

The choice of detector for non-chromophoric samples in SFC istraditionally limited to Flame-ionization detection (FID). However,the use of FID is not possible when organic modifiers are present.ELSD is ideally suited to the technique of SFC and to the analysisof non-chromophoric samples using SFC. Oligomeric separationof medium to high molecular weight PEGs by SFC is considerablyfaster than by reversed phase HPLC.

Samples: PEG 400, PEG 600 & PEG 1000Column: Commercially available Diol Column 60Å 6µm, 250x4.6mmEluent : Carbon Dioxide with 5% Methanol Flow Rate: 2.0ml/minPressure: Carbon Dioxide was pressure programmed

from 100bar to 250bar, at 10bar/min. Inj Vol: 10µlDetector: PL-ELS 2100 (neb=40°C,

evap=30°C, gas=0.8 SLM)

400 min

1

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Key Applications of the PL-ELS 2100 Evaporative Light Scatte

KEY1. GGG2. LGG3. LGL4. LLL5. OOO

440 min

200 min

1

23

Sample: 2mg/ml Borage OilColumn: Commercially availabEluent A: ACNEluent B: DCMGradient: 30-50% B in 40 mins,

hold 3 minsFlow Rate: 1.0ml/minInj Vol: 20µlDetector: PL-ELS 2100 (neb=25

gas=1.4 SLM)

5

4

3

440 min

440 min

Polymer AdditivesColumn: Commercially available C18 5µm, 150x2.1mm (40°C)Eluent A: 0.1% TEA in WaterEluent B: ACNGradient: 70-100% B in 10 mins,

hold 8 minsFlow Rate: 0.4ml/minInj Vol: 20µlSample Conc: 100µg/ml eachDetector: PL-ELS 2100 (neb=40°C,

evap=85°C, gas=1.1 SLM)

KEY1. Ultranox 6262. Irganox 10103. Ethanox 3304. Alkanox 2405. Erucamide

250 min

1

2

3

4

5

PEG 400

PEG 1000

PEG 600

Sugars in Aloe JuiceStudies in the field of glycomics suggest that the monosaccharidecontent in aloe juice contributes significantly towards its anti-inflammatory activity.

Column: PL Hi-Plex Ca, 300x7.7mmEluent: WaterFlow Rate: 0.6ml/minInj Vol: 20µlSample Conc: 1mg/mlTemp: 80°CDetector: PL-ELS 2100 (neb=50°C, evap=90°C, gas=1.6 SLM)

KEY1. Trehalose2. Glucose3. Fructose

■ The ELSD offers significant improvements over RI detection for the analysis of sugars such as glucose (monosaccharide) and trehalose (disaccharide), as it has increased sensitivity and better baseline stability

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Efficient Evaporation of Complex EluentsResolution of phospholipids can be difficult to achieve withoutusing a complex eluent gradient. PL’s evaporative light scatteringdetectors can efficiently evaporate even the most complexgradients, providing ideal detection of polar lipids.

Samples: 1. Cholesterol 2. Phosphatidylethanolamine (PE), 3. Phosphatidylcholine (PC), 4. Sphingomyelin (Sph), 5. Lysophosphatidylcholine (LPC)

Column: Commercially available DIOL column, 150x2.1mmEluent A: IPA/Hexane/Water/Ammonia Hydroxide (57.8/40/2/0.2)Eluent B: IPA/Hexane/Water/Ammonia Hydroxide (51.8/40/8/0.2)Gradient: 10-100% B in 7 mins (hold 8 mins), 100-0% B in 5 mins

(hold 10 mins)Flow Rate: 0.3ml/minInj Vol: 10µlDetector: PL-ELS 2100 (neb=30°C, evap=80°C, gas=0.8 SLM)

■ Complex eluents are easily removed with the PL-ELS 2100

120 min

1

23

4 5

Triglycerides in Cooking OilThe triglycerides composition of cooking oil is very complex andcontains many chemically similar species. In order to develop ahigh resolution HPLC separation of these components, gradientelution is required, normally involving solvents with very differentproperties. For this reason, RI is not viable as a detection methodand UV can be difficult due to baseline disturbance arising fromthe choice of solvents used in the gradient.

Sample: Sesame OilColumn: Commercially available C18 5µm,

250x4.6mmEluent A: ACNEluent B: DCMGradient: 30-50% B in 40 mins,

50-90% B in 2 mins, hold 3 minsFlow Rate: 1.0ml/minInj Vol: 20µlDetector: PL-ELS 2100 (neb=25°C,

evap=50°C, gas=1.4 SLM)

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Underivatized Amino AcidsThere are numerous methods involving gas and liquidchromatography coupled with pre- and post- columnderivatization for the analysis of amino acids. These methods arewell established and can offer low detection limits, however, theyare time consuming and the products resulting from thederivatization step can be unstable.

Column: Commercially available ODS 5µm, 250x4.6mmEluent A: 0.1% TFA in WaterEluent B: 0.1% TFA in ACNGradient: 100% A, 5 min hold, 0-40% B in 15 minsFlow Rate: 1.0ml/minInj Vol: 10µlSample Conc: 1mMDetector: PL-ELS 2100 (neb=50°C, evap=50°C, gas=1.6 SLM)

KEY

1. Serine2. Glutamic acid3. Arginine4. Proline5. Valine6. Methionine7. Isoleucine8. Leucine9. Phenylanine10. Tryptophan

■ ELSD is ideal for the detection of amino acids as no derivatization step is required, thus reducing the time and cost for sample preparation and analysis

300 min

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30 min

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ering Detector

200 min

1

2le C18 5µm, 250x4.6mm

50-90% B in 2 mins,

5°C, evap=50°C, 5

43

400 min

1

2

3

4

5

6

7

8

910

KEY1. Cholesterol 2. Phosphatidylethanolamine3. Phosphatidylcholine (PC)4. Sphingomyelin (Sph)5. Lysophosphatidylcholine (LPC)

AcetaminophenAcetaminophen belongs to classes of drugs called analgesics(pain relievers) and antipyretics (fever reducers), and since it wasapproved by the FDA in 1951, has been used in many over thecounter medication brands. ELSD permits the determination oflow levels of acetaminophen; shown here is a chromatogram foran on column loading of 2µg.

Column: Commercially available C18 5µm, 150x4.6mm

Eluent A: 90% Water : 9% MeOH : 1% Acetic acid

Eluent B: ACNIsocratic: 60/40% A/BFlow Rate: 1.5ml/minInj Vol: 10µlSample Conc: 200µg/mlDetector: PL-ELS 2100

(neb=25°C, evap=25°C, gas=1.6 SLM)

■ The PL-ELS 2100 provides excellent sensitivity for the analysis of semi-volatiles at room temperature

■ This close-up illustrates the excellent peak shape of the PL-ELS 2100 which gives high resolution of complex samples

KEY1. LLL2. LLO3. LLP4. OOL5. POL6. PPL7. OOO8. OOP9. PPO10. OOS

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Sub-ambient ELSD for Low Boiling Point Compounds

Low temperature ELSD is essential when analyzing thermallylabile compounds. Current ELS detectors cannot operate lowerthan ambient temperature (i.e. 25-30°C), but even at ambienttemperature, the response of low molecular weight (e.g. 60-150MW) semi-volatile compounds by ELS detection can be poor.

To improve the detection of thermally unstable compounds,Polymer Laboratories has pushed the boundaries of ELSdetection with the PL-ELS 2100 Ice. Using the same evaporatorgas technology seen in the PL-ELS 2100, the PL-ELS 2100 Icecan be operated at temperatures as low as 10°C.

Operating at sub-ambient temperatures provides maximumsensitivity for semi-volatile compounds that show poorresponses at ambient temperature. The advantage of using thePL-ELS 2100 Ice is shown opposite for a mixture ofpharmaceutical compounds.

Pharmaceutical Analysis at Sub-ambient Temperature

When analyzing compound mixtures of equal concentration,evaporative light scattering detectors can show distinctdifferences in response between compounds. Most of the time,these variations can be attributed to differences in thecompounds’ volatility.

These volatility differences are emphasised at higher ELSDtemperatures (e.g. 50°C), as shown in the figure below. In order tominimize or eliminate these volatility effects, the operatingtemperature of the ELSD needs to be as low as possible.

By operating the PL-ELS 2100 Ice at sub-ambient temperatures,the volatility differences observed at 50°C are eliminated at 15°C,producing a uniform response (see below).

Column: Commercially available C18 5µm, 150x4.6mmEluent A: Water + 0.1% TFAEluent B: ACN + 0.1% TFAGradient: 60-90% B in 5 minsFlow Rate: 1.0µl/minInj Vol: 20µlDetector: PL-ELS 2100 Ice

(gas=1.6 SLM)

KEY1. Acetanilide2. Indapamide3. Ibuprofen4. Dibutylphthalate

80 min

24°C

35°C

15°C

1

3

4

1

2

3

4

1 2

3

4

2

NEWPL-ELS 2100 Ice (Integrated Cooled Evaporator)

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4.50 min

S/N = 350

S/N = 64

20oCgas=1.85 SLM

S/N = 9.0

25oCgas=1.4 SLM

30oCgas=1.4 SLM

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Hydrocarbon AnalysisEven compounds with high boiling points can benefit from the PL-ELS 2100 Ice, as shown below. The hydrocarbons,hexadecane (Bp 287°C), heptadecane (Bp 302°C) and eicosane(Bp 343°C) have high boiling points that suggest they are easilydetectable by ELSD. However, even at 30°C, there is a distinctdisparity in response between the three hydrocarbons. Operatingthe PL-ELS 2100 Ice at 18°C dramatically improves the responseof hexadecane and heptadecane, such that they saturate thedetector signal.

Sample: Aliphatic Hydrocarbons, 0.2mg/ml Eluent: THFFlow Rate: 1.0ml/minInj Vol: 10µl 30 secs apart on PLgel guard columnDetector: PL-ELS 2100 Ice (neb= 80°C, gas=1.1 SLM)

KEY1. Hexadecane (C16)2. Heptadecane (C17)3. Eicosane (C20)

70 min

20oC

18oC

30oC1 2

3

30 min

S/N 61

S/N 15

30oC

15oC

Sample: PEG 106, 1mg/ml Column: PLRP-S 100Å 5µm, 150x4.6mmEluent: 10% ACN in WaterFlow Rate: 1.0ml/minInj Vol: 10µlDetector: PL-ELS 2100 Ice

(neb= 30°C, gas=varied)

■ LoD Improved ca. 40 Fold !!!

For higher throughput of samples, uniquely, the PL-ELS 2100 Ice features rapid temperatureequilibration; the detector can cool down from 50°C to30°C, and vice versa, in just over five minutes.

0 4321 5

50

30

35

40

45

min

Tem

p °C

Improve the Limit of Detection for Low MolecularWeight CompoundsThe limit of detection for thermally sensitive analytes, such asacetanilide (MW 135.2) and Polyethylene Glycol (MW 106), is oftenlimited by their volatility. Using the PL-ELS 2100 Ice, the detectionlimit for these types of compounds can be dramatically improved.

Sample: 200μg/ml AcetanilideColumn: Commercially available C18 5µm, 150x4.6mmEluent A: Water + 0.1% TFAEluent B: ACN + 0.1% TFA Gradient: 60-90% B in 5 minsFlow Rate: 1.0ml/minInj Vol: 20μlDetector : PL-ELS 2100 Ice

(neb= 30°C, gas=1.6 SLM)

■ LoD Improved 4 Fold !!!

■ In addition to the enhanced detection of semi-volatile compounds, the temperature controlled evaporator tube on the PL-ELS 2100 Ice allows the user to rapidly swap between temperature programs

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PL-ELS 1000 - For HPLC and GPC Applications, Ambient to High

The PL-ELS 1000 For HPLC and GPC Applications,Ambient to High Temperature

The PL-ELS 1000 is a wellestablished market leaderrenowned for its ruggeddesign and able to deliverhigh performance, even forvery demanding HPLC orGPC applications. Thenebulizer and evaporator canbe controlled at very hightemperatures to efficientlyhandle high boiling pointsolvents commonly used inGPC that other ELSDs simplycannot manage.

A custom designed heated transfer line, which is also controlledfrom the PL-ELS 1000, permits easy coupling to any GPC system,especially valuable for use in high temperature GPC applications.

The solvent condensationdevice allows the recovery ofmaterial from the nebulizerdrain and the solvent vaporfrom the exhaust forincreased safety, particularlyimportant when using morehazardous solvents.

The PL-ELS 1000 advanced control software is extremely easy touse, providing temperature and gas flow control, signalmonitoring, methods storage, system automation and logging.

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1 2

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min0

Fatty AcidsELSD is ideal for the analysis of both saturated and unsaturatedfatty acids, as these compounds have no UV chromophore andrequire gradient elution.

Column: PLRP-S 100Å 5µm, 250x4.6mmEluent A: 60mM Acetic acidEluent B: ACNEluent C: THFGradient: 35/60/5 A/B/C to 0/90/10 A/B/C in 20 minsFlow Rate: 0.5ml/minTemp: 45°CDetector: PL-ELS 1000 (neb=80°C, evap=70°C, gas=1.0 SLM)

KEY1. Capric acid2. Lauric acid3. Myristic acid4. Palmitic acid5. Stearic acid

■ Working at low pH to maximize resolution and minimize analysis time, the ELSD provides good sensitivity and a stable, drift free baseline

210 min

210 min

1

2

3

RI

1

2

3

PL-ELS 1000

Analysis of Polymer Blends and CopolymersPolymer blends often combine homopolymers with widely differingcharacteristics. The PL-ELS 1000 gives a positive response for allsamples regardless of chemistry, and is the ideal detector foranalysing polymer blends by GPC, as illustrated by thecomparison of the RI and PL-ELS 1000 below.

Sample: Blend of Polystyrene (PS) and Polydimethylsiloxane (PDMS)Columns: 2xPLgel 5µm MIXED-C, 300x7.5mmEluent: TolueneFlow Rate: 1.0ml/minDetectors: PL-ELS 1000 (neb=100°C, evap=120°C, gas=1.5 SLM)

RI

KEY1. PS2. PDMS3. Blend

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High Temperature GPCPolyolefins are typically analyzed by GPC using trichlorobenzene(TCB) at 160°C. As the dn/dc of polyolefins and polystyrenestandards in TCB is relatively small, RI detector performance mustbe maximized to obtain acceptable signal to noise ratio. Thepolystyrene response in TCB using the PL-ELS 1000 is greatlyimproved compared to a typical RI chromatogram, illustrating theincreased sensitivity of the PL-ELS 1000 for high temperatureGPC applications.

Sample: PolyethyleneColumns: 2xPLgel 10µm MIXED-B, 300x7.5mmEluent: TCBFlow Rate: 1.0ml/minInj Vol: 200µlSample Conc: 1mg/mlTemp: 160°CDetector: PL-ELS 1000 (neb=160°C, evap=270°C, gas=1.5 SLM)

KEY1. 2,560,0002. 320,0003. 59,5004. 10,8505. 580

■ The PL-ELS 1000 is the ONLY ELSD on the market which operates at this temperature

11

www.polymerlabs.com/elsd

h Temperature

0%

25%

15%

35%

50%

20min1

205 min

1

23 4

5

PL-ELS 1000

RI

203 min

Polymer MW Analysis using GPC/SECWith conventional RI detectors, ambient temperature effects,outgassing with low boiling solvents and simple eluentreplenishment, can lead to drifting and noisy baselines. The PL-ELS detectors have no flow cell and therefore theseproblems are eliminated as the solvent is evaporated (and playsno part in the detection). In addition, the PL-ELS 1000 provides adramatic increase in sensitivity to allow a reduction in columnloading - key for some high molecular weight samples.

Sample: Poly(lactide-co-glycolide)Columns: 2xPLgel 5µm MIXED-D, 300x7.5mmEluent: ChloroformFlow Rate: 1.0ml/minInj Vol: 20µlSample Conc: 2mg/mlDetector: PL-ELS 1000 (neb=50°C,

evap=70°C, gas=1.0 SLM)

■ The PL-ELS 1000 also offers a significant increase in sensitivity, by a factor of 50

Copolymer Characterization by GPECCopolymers are often characterized for composition usinginteractive HPLC methods, usually employing gradient elutionwith a wide variety of solvents depending on the application. The PL-ELS 1000 responds to all polymers, irrespective of theirchemical composition, with no deterioration in baseline stability,whatever the choice of gradient.

Sample: Random poly(lactide-co-glycolide) polymers with varying glycolide content

Column: PLRP-S 1000Å 8µm, 150x4.6mmEluent A: MeOHEluent B: THFGradient: 2 min hold in 99% A, 1-99% B in 10 minsFlow Rate: 1.0ml/minDetectors: PL-ELS 1000 (neb=70°C, evap=80°C, gas=1.0 SLM)

% Glycolide content

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www.polymerlabs.com/elsd

Item Part No.

PL-ELS 2100 (110V) PL0860-0110PL-ELS 2100 (240V) PL0860-0240

PL-ELS 2100 Ice (110V) PL0860-1110PL-ELS 2100 Ice (240V) PL0860-1240

PL-ELS 1000 (110V) PL091-24797-110PL-ELS 1000 (240V) PL091-24797-220

Your local distributor is:

Contact the UK office for details of your local distributor

All Trademarks Acknowledged Polymer Laboratories reserves the right to change specifications without notice.

Specifications

North AmericaPolymer Laboratories Varian, Inc.Amherst Fields Research Park160 Old Farm Road, Amherst MA 01002, USATel, toll free 800 767 3963or tel 413 253 9554Fax 413 253 2476Email [email protected] [email protected] Cards Accepted.

BeneluxPolymer Laboratories BVSourethweg 1, 6422 PC HeerlenThe NetherlandsTel (+31) 045 5414748Fax (+31) 045 5410005Email [email protected]

New Benelux Address - From April 2006Polymer Laboratories BVHerculesweg 8, 4338 PL MiddelburgThe NetherlandsTel (+31) 011 8671500Fax (+31) 011 8623193

Now a part of Varian, Inc.

UK / InternationalPolymer Laboratories LtdEssex Road, Church StrettonShropshire SY6 6AX, UKTel (+44) 01694 723581Fax (+44) 01694 722171Email [email protected] [email protected] Cards Accepted.

FrancePolymer Laboratories SARLGVIO Parc de Marseille SudImpasse du Paradou Bâtiment D5BP 159 13276 Marseille Cedex 09, FranceTel (+33) 04 91 17 64 00 Fax (+33) 04 91 17 64 01 Email [email protected] www.polymerlabs.fr

GermanyPolymer Laboratories GmbHPEKA Park T5 (001), Otto-Hesse Straße 19D-64293 Darmstadt, GermanyTel (+49) 06151 860690Fax (+49) 06151 860670Email [email protected]

US Patent No. 6,151,113 and foreignequivalents and UK Patent ApplicationNo. 0304253.8 apply

Ordering Information

CS

804/

B/0

2.06

PL-ELS 2100 PL-ELS 2100 Ice PL-ELS 1000Light Source LED 480nm LED 480nm Tungsten/Halogen lamp

Detector Photomultiplier tube Photomultiplier tube Photodiode

Temperature RangeEvaporator Ambient to 120°C 10 - 80°C 30 - 300°C Nebulizer Ambient to 90°C Ambient to 90°C 30 - 220°C Heated Transfer Line (optional) Not available Not available 30 - 220°C

Nebulizer GasTypical Inlet Pressure 60 - 100psi 60 - 100psi 60 - 100psi Maximum Inlet Pressure 100psi 100psi 150psi Gas Flow Rate 0 - 3.25 SLM 0 - 3.25 SLM 0 - 1.0 SLM

Eluent Flow Rate 0 - 5ml/min 0 - 5ml/min 0 - 0.5ml/min

Analog Output 0 - 1V FSD 0 - 1V FSD 0 - 1V and 0 - 10V FSD

Digital Output 24 bit (10Hz) via serial port 24 bit (10Hz) via serial port

Communications Serial I/O (RS232), Serial I/O (RS232), Serial I/O (RS232),contact closure, contact closure, contact closure, TTL TTL TTL, heated transfer line control

Operation Graphical Vacuum Fluorescent Graphical Vacuum Fluorescent Detachable IR control pad ordisplay and keypad or Windows display and keypad or Windows Windows based PC control based PC control based PC control

Power Requirements 90/120V AC or 220/250V AC, 90/120V AC or 220/250V AC, 90/120V AC or 220/250V AC, 50/60Hz 50/60Hz 50/60Hz

Dimensions (wxdxh) 200x450x415mm 200x450x415mm 175x480x430mm

Weight 11 kg 13 kg 15 kg

Now a part of Varian, Inc.