Detect Everything You Inject With the PL-ELS 2100 Evaporative Light Scattering Detector

First Impressions Compact, modern design Flat top Small footprint, stackable Operation via control panel and simple keypad Column connection at front of detector One exhaust tube at rear, one liquid waste port at front

PL-ELS 2100 Front Panel Detail 200mm 200mm Display Keypad 415mm Column Connection Liquid Waste 450mm 450mm

Principle of Operation The ELSD principle of operation employs three distinct stages : Nebulization Evaporation Detection Gas Eluent Inlet Light Source Liquid Waste

Nebulization Eluent flow mixed with N2/Air Concentric nebulizer gives efficient nebulization, stable droplet plume, uniform droplet size Narrower cone of spray plume allows a smaller nebulizer chamber, which minimizes band broadening Better recovery of semi-volatile solutes compared to cross-flow nebulizer Easier to align, reproducible from one detector to another Temperature independently controlled

Evaporation Solvent removed by heating Temperature independently controlled PL-ELS 2100 has an additional gas input that aids evaporation (patent pending) Small volume evaporator Fast thermal equilibration Diffuser reduces turbulence inside evaporator tube which minimizes band broadening (patented)

Detection Blue LED light source (480nm) Intensity controllable by user Photomultiplier detector tuned to same wavelength for increased sensitivity Feedback control for light source provides constant output Detector mounted directly onto the optics which eliminates stray light loss and increases sensitivity Reduced volume in the optics chamber, minimizes band broadening Digital gain control to optimize full scale sensitivity Digital signal smoothing to optimize S/N ratio

Advantages of Evaporative Light Scattering Detection Universal: responds to all compounds in the mobile phase. - Detects compounds that do not possess a UV chromophore e.g. polymers, sugars

Advantages of Evaporative Light Scattering Detection: Pharmaceutical Mixture Detects all components in a single run Samples : 1. α-cyclodextrin 2. ß-cyclodextrin 3. Ibuprofen Column: Aquasil C18 5µm, 150x4.6mm Eluent A: Water Eluent B: Acetonitrile Gradient: 50-95% B in 5 mins Flow Rate: 1.0 ml/min Inj Vol: 20µl Detector: PL-ELS 2100 (neb=30°C , evap=50°C, gas=1.0 L/min) ELSD UV @ 220nm

Advantages of Evaporative Light Scattering Detection Universal: responds to all compounds in the mobile phase - Detects compounds that do not possess a UV chromophore e.g. polymers, sugars Not dependent on spectroscopic properties of compound: - Produces more uniform detection sensitivity for analytes

Advantages of Evaporative Light Scattering Detection: Uniformity of Response Comparison of UV and ELS traces show that ELSD allows for a more uniform response than UV detection Column: PLRP-S 100Å 5µm, 150 x4.6mm Eluent A: 50% 0.1% TFA in Water : 50% 0.1% TFA in ACN Flow Rate: 1.0ml/min Inj Vol: 10µl Detector: PL-ELS 2100 (neb=30°C, evap=30°C, gas=1.4 L/min) UV-VIS @ 280nm

Advantages of Evaporative Light Scattering Detection Universal: responds to all compounds in the mobile phase Detects compounds that do not possess a UV chromophore e.g. polymers, sugars Not dependent on spectroscopic properties of analyte: - Produces more uniform detection sensitivity for analytes Not susceptible to baseline drift during gradient elution, temperature or solvent pump fluctuations ELSD compatible with a much wider range of solvents compared to RI and UV

Fast Gradient, Fast Flow Rate Capability Sample: Indapamide (IND), Dibutyl phthalate (DBP) Column: PLRP-S 100Å 5µm, 50x4.6mm Eluent A: 0.05% TFA in Water Eluent B: 0.05% TFA in ACN Gradient: 5-95% B in 1 min Flow Rate: Increased from 2ml/min up to 5ml/min Detector: PL-ELS 2100 (neb=30°C, evap=30°C, gas=1.6 L/min) Note : IND is non-volatile, DBP is relatively volatile

Fast Gradient, Fast Flow Rate Capability 2ml/min 3ml/min 4ml/min 5ml/min IND DBP min

Fast Gradient, Fast Flow Rate Capability Stable baseline throughout the gradient PL-ELS 2100 can operate in 100% up to 5ml/min 2ml/min 3ml/min 4ml/min 5ml/min min

Similar operating principles to LC-MS Advantages of Evaporative Light Scattering Detection: Ideal Complement to LC-MS Similar operating principles to LC-MS - Volatile buffers - Favors lower flow rates (i.e. 0.2-0.5 ml/min) Can develop LC methods on PL-ELS 2100 then transfer to LC-MS ELSD can provide supporting information when used in tandem with LC-MS

Ideal Complement to LC-MS Sample Mixture of known 1:1 ratio LC-MS results show ratio to be 3:1 UV-Vis result show ratio to be 10:1 PL-ELS 2100 results show ratio to be 1:1 (Response independent of optical properties)

PL-ELS 2100 Sensitivity - Limit of Detection Column: C18 5µm,150x4.6mm Eluent A: 99% Water + 0.1 % TFA Eluent B: 1% ACN + 0.1% TFA Flow Rate: 0.5ml/min Detector: PL-ELS 2100 (neb=50°C, evap=70°C, gas=1.2 L/min) Sample: DL-Glutamic Acid

PL-ELS 2100 Sensitivity - Limit of Detection LOD improved with lower eluent flow rates and smaller ID column Caffeine Loading 100ng 4.6 ID S/N = 4.4 Caffeine Loading 30ng 2.1 ID S/N = 11.5

Operating the PL-ELS 2100: Onboard Instrument Control Interactive menu bar Current method Mode Detector output Status Actual values of set parameters Keypad

Two modes of operation : Simple Operation Two modes of operation : STANDBY Power on LED off Heaters off Gas at minimum flow rate (1.2 L/min) - Gas shuts off after 15 minutes RUN LED on Temperatures and gas flow controlled to set values

Digital Signal Processing Digital Signal functions are accessed via the down arrow key in main menu screen Parameter Screen allows you to change - PMT output or gain (1-250) - Smoothing (1-50)  - LED intensity (0-100)

PL-ELS 2100 Control Software Select mode of operation Control parameters Evaporator & Nebulizer temp Gas flow (0-3.25 L/min) Monitor output Gain setting (0.1-250) Signal smoothing (0-5) Log instrument conditions over time Load / Edit methods - 10 programmable methods

PL-ELS 2100 Control Software Supplied as standard with the PL-ELS 2100 Select 1 of 10 different temperature programs according to analysis Also available as a Software Development Kit for third party system integration To use the control software simply connect the PL-ELS 2100 to a PC via a serial connection (RS232) on selectable COM port Where no serial port is available, a USB serial connector can be used

PL-ELS 2100 Method Manager Software Used when the instrument is to be controlled via the onboard instrument control (rather than software control) This is a simple software utility designed to set up, edit and log defined methods which can then be downloaded to the PL-ELS 2100 Method Manager Software supplied as standard with the To use this software utility, simply connect the PL-ELS 2100 to a PC via a serial connection (RS232)

PL-ELS 2100 Method Manager

PL-ELS 2100: Routine Operation When running with the “XXX” method, the User has complete flexibility to change parameters via the PL-ELS 2100 keypad and display (e.g. for method development) Once standard methods for operation of the PL-ELS 2100 have been established, these can be stored using the PL-ELS 2100 Method Manager software utility and downloaded to the unit These downloaded parameters are stored as numbered methods (1-10) and cannot be edited through the keypad, only by running the PL-ELS 2100 Method Manager software

A New Approach to ELS Detection Operating conditions of the PL-ELS 1000 is based on the properties of the eluent (e.g. high boiling point solvents require high evaporator temperatures) However, the PL-ELS 2100 operating conditions are determined by the properties of the analyte (e.g. semi-volatile or non-volatile) The PL-ELS 2100 can evaporate water at ambient temperature, unlike the PL-ELS 1000.

Manipulating the PL-ELS 2100 Parameters FOR NON-VOLATILE SOLUTES Nebulizer and evaporator can run hot to increase signal to noise ratio Gas flow is set as low as possible to maximize signal FOR RELATIVELY VOLATILE SOLUTES Nebulizer and evaporator run cold to increase signal Gas flow is increased to control baseline noise

Volatile Applications: Effect of Changing Evaporator Temperature Samples: 1. Acetanilide, 2. Indapamide, 3. Ibuprofen, 4. Dibutylphthalate Column: C18 5µm, 150x4.6mm Eluent A: 0.1% TFA in Water Eluent B: 0.1% TFA in ACN Gradient: 60-90%B in 5 mins Flow Rate: 1.0mL/min Detector: PL-ELS 2100 (neb and evap set at the same temperature, gas 1.8 L/min) Note: peak 2 is non-volatile, peaks 1, 3, 4 are relatively volatile

Volatile Applications: Effect of Changing Evaporator Temperature At high temperature peak 2 has better S/N but peaks 1,3,4 are not detected At 30°C all four peaks are detected

Environmental Application:Herbicides Sample: Mixture of 10 Phenylurea herbicides Column: C18 5µm, 250x4.6mm Eluent A: Water Eluent B: Acetonitrile Gradient: 10-80%B in 40 mins Flow Rate: 0.7ml/min Inj Vol: 20µl Detector: PL-ELS 2100 (neb=25°C, evap & gas flow varied)

Environmental Application: Herbicides

Non-Volatile Applications: PEG 400 Separation Sample: PEG 400 Column: PLRP-S 100Å 5µm, 150x4.6mm Eluent A: Water Eluent B: ACN Gradient: 10-30% B in 12 mins Flow Rate: 1.0mL/min Inj Vol: 10µl

PEG 400: Effect of Changing Temperature Higher temperatures increase S/N, no risk of loss due to evaporation Evap 90°C Neb 90°C Gas 1.6 L/min Evap 30°C Neb 30°C Gas 1.6 L/min

PEG 400: Effect of Changing Gas Flow Sensitivity is improved as gas flow is reduced Evap 70°C Neb 50°C Gas 1.2 L/min Evap 70°C Neb 50°C Gas 2.4 L/min

Non-Volatile Application: Triglycerides in Natural Oils Sample: 2mg/ml Starflower Oil Column: C18 5µm, 250x4.6mm Eluent A: ACN Eluent B: DCM Gradient: 30-50% B in 40 mins; 50-90% in 2 mins, hold for 3mins Flow Rate: 1.0ml/min Inj Vol: 20µl Detector: PL-ELS 2100 (neb=40°C, evap=70°C, gas=1.4 L/min)

Non-Volatile Application: Triglycerides in Natural Oils

Supercritical Fluid Chromatography: PEG Separation Sample: PEG 1000 Column: Berger SFC Diol Column 60Å 6µm, 250x4.6mm Eluent: Carbon Dioxide with 5% Methanol Flow Rate: 2ml/min, Pressure: Carbon Dioxide was pressure programmed from 100bar to 250bar at 10bar/min. Inj Vol: 10µl Detector: PL-ELS 2100 (neb=40°C, evap=30°C, gas=0.9 L/min)

Supercritical Fluid Chromatography PEG Separation Reduced band broadening allows high resolution SFC separations to be performed PEG 1000

PL-ELS 2100: DMSO Transparency Sample: Pharmaceutical Mixture in DMSO Column: C8 5µm, 50x4.6mm Eluent A: Water Eluent B: Acetonitrile Gradient: 5-100% B in 5 mins Flow Rate: 1.0ml/min Inj Vol: 20µl Detector: PL-ELS 2100 (neb=25°C, evap & gas flow varied)

PL-ELS 2100 DMSO Transparency: Ideal for High Throughput Screening Loss of volatile species occurs as temperature is increased

PL-ELS 2100: DMSO Transparency: Ideal for High Throughput Screening The design of the PL-ELS 2100 makes it transparent to DMSO at 25-30°C Great potential in the High Throughput Combichem market Unique to the PL-ELS 2100 due to its use of evaporation gas PL-ELS 2100: neb=25°C, evap=30°C

Benefits of Evaporative Light Scattering Detection Universal detector: independent of the optical properties of the solute Compatible with gradient elution Ideal for fast gradient elution:- no solvent front. Removes the need for derivatization Compatible with chromatographic techniques such as MS and SFC Wide application area

Additional Benefits of the PL-ELS 2100 Operation in 100% water at 25°C DMSO transparency at ambient temperature Extremely low dispersion for high resolution separations (e.g. SFC) High sensitivity to volatile compounds Compatible with eluent flow rates, up to 5ml/min Rapid thermal equilibration Easy to use Extremely small footprint, stackable