Firma:                   Thermo Separation

Modell:                CE-Ultra

Kommentar:        Dokumente engl.

Introduction

Capillary electrophoresis (CE) is the movement of ions through a capillary subjected to an electrical field. Sample ions are introduced hydrodynamically (using pressure differences) or electrokinetically (using charge) to draw sample into a capillary. Once the sample is injected, the end of the capillary is removed from the sample reservoir and immersed in the running buffer. Separation is based an the charge-to-mass ratlos of the constituent ions. As the sample migrates through the capillary, a variable-wavelength detector monitors the presence or absence of the constituents of interest. The detection data is then sent to a data acquisition system, which creates a recording of the electrophoretic separation, an electropherogram or E-gram ^TM , of the run.

SpectraPHORESIS ULTRA System Operation

A typical automated CE system contains seven functional components: 1) fused-silica capillary, 2) electrolyte vessels, 3) electrodes, 4) pressure system. 5) regulated high-voltalte power supply, 6) detector, and 7) data acquisition system. The following Instrument modules comprise the SpectraPHORESIS ULTRA System:

SpectraPHORESIS VialServer (Autosampler): The VialServer holds up to 110 sample or buffer vials in peeparation for CE analyses. The optional Tray Temperature Control function maintains up to 95 vials at temperatures from 4° - 60°C. The vial positions can be prearranged and easily coordinated with the acquisition method defined in the PC1000 System Software. The VialServer's Sample/Buffer Preparation function allows for automated pre-run huffer preparation and post-run huffer replenishment.

SpectraPHORESIS ULTRA (CE Module): Electrophoretic separation occurs in the CE module. The CE module shuttle automatically slides into the side access door in the VialServer to ohtain buffer and sample vials as specified in the sequence method. The shuttle then slides hack into the CE module to transport the required vials to the front and rear vial positions in the injection tower. There they are raised onto the ends of the fused-silica capillary housed in the capillary cassette. The cassette is inserted into the capillary oven, which controls the capillary temperature throughout the run.

The capillary is threaded between two ball lenses. Prior to injection, and as the vial is lifted into the injection tower, the vial septa are pierced by the needle. The vials are then raised into the final pre­injection position, causing the electrode, and capillary to protrude through the vial septa and become immersed in the vial contents. The injection tower needle septa combined with vial and cassette septa work together to maintain a stable injection pressure, minimizing backflushing into the needle and loss of precious sample. This unique tri-seal configuration contributes to the 2% RSD precision offered by the CE ULTRA System. During hydrodynamic injection, the positive­pressure pneumatic system forces sample into the capillary. During electrokinetic injection, the high-voltage power supply provides potential up to 30kV to draw the sample into the capillary.

SpectraPHORESIS UV3000: Separated sample constituents pass through the dual-ball sapphire lens assembly located in the lower portion of the cassette. Either deuterium (UV 190-365 nm) or tungsten (visible 366-800 nm) light emerging from the capillary is sent to the sample diode in the SpectraPHORESIS UV3000 scanning detector. A reference signal is simultaneously sent to the detector reference photodiode. The detector can run in single, dual, triple wavelength, or scanning modes.

PC1000 Software: The SpectraPHORESIS ULTRA modules are connected to a personal Computer where instrument control, acquisition and calculation method setup, sequence and vial assignment, data acquisition, data processing, and reporting are Bone using Thermo Separation Products" PC1000 Software Ver 3.5, or higher. Refer to the PC1000 System Software Reference Manual shipped with your SpectraPHORESIS ULTRA System for further information an PC1000 software.

PC 1000 processes the data acquired from the SpectraPHORESIS UV3000 to perform peak detection and quantification. The software also provides real-time Status information throughout the run and can create a post-run report and an electropherogram of the Separation. Post-run reprocessing of previously acquired data is also possible for further method refinement.

Specifications

Safety Features:                      Interlock disabling of high-voltage CE module

Power Requirements:             100/120/240 VAC nominal. 50/60 Hz

Ambient Environment:             10 to 40 °C; 5%-95% RH non-condensing

Bench Space without PC:       44 x 92 52 cm (H x W x D) cm total bench spare without computer

SpectraPHORESIS VialServer

Maximum Vial Capacity:          110 vials, 95 with optional tray thermostatting

Tray Temperature Control:     Optional 0-60 °C, Peltier controlled

Needle Height:                         Programmable in 0.1 mm increments

Vial Dimensions/Volume:        35 x 11.5 mm. Blass or plastic (ca. 1.8 mL glass, ca 650 µL and 100 µL plastic, screw capped. 100 µL. glass microinserts).

Sample/Buffer Preparation

Reagent Vials:                         Up to three vials per method

Mixing/Heating:                        Single vial. air-driven vortex heater/mixer. settahle in 1°C increments form 30° to 100 °C

Dilution Precision:                   Typically <1.0% RSD for 1/100 dilution (10 µL to 1 rnL)

Dispensing Precision:             Typically <0.2% RSD for 1 mL dispensing from external solvent reservoir

Dimensions and Weight:        37 x 41 x 51 cm (H x W D); 16 kg Standard. 24 kg with all options installed

SpectraPHORESIS ULTRA (CE Module)

Vial Shuttle Capacity:              Up to 2 x 5 vials

Minimum Sample Volume:      20 µL pressure injection. 50 µL electromigration sampling (using microinserts)

Wash                                       100 psi pressure, applicable to either of the two capillary ends

Capillary Temperature Control:

Peltier controlled, 10°C to 60°C in 0.1°C increments. Maximum ringe of cooling 10°C below amhient. Modes: constant temperature. temperature gradients, constant resistance

Run Modes:

Constant voltage, constant current, voltage gradients, positive or negative polarity, time-programmed switches. Apply pressure at one or both ends of a capillary with the voltage or current applied.

Capillary Cassette:                  Bar code labeled. Minimum capillary length ca. 20 cm. Maximum length 100 cm. Shortest detection length (Ld) 5 cm.

Dimensions and Weight:        44 x 36 x 51 cm (H x W x D); 27 kg.

SpectraPHORESIS UV3000 (Detector)

Modes:                                                Time-programmable single wavelength, multiple wavelengths. scanning

Wavelength Range:                190 - 800 nm

Wavelength Accuracy:            ± 1 nm

Wavelength Precision:            0.1 nm

Lamps:                                    Deuterium and tungsten, pre-aligned

Optics,:                                    Fiber optics. ball lenses

Rise Time:                               0.0 to 9.9 sec

Noise:                                      2.5 x 10 ^-5 AU @ 210 nm

Dimensions and Weight:        37 x 15 x 47 cm (H x W x D); 11 kg

PC1000 System Requirements

COMPUTER HARDWARE

Refer to the TSP README file for the specific system requirements. These include

CPU                                         486 66MHZ or higher

Monitor                                     SVGA or higher

Math Coprocessor                  Required

Memory                                   16 Mbytes (minimum), 32 MB strongly recommended

Disk Space                              10 Mbytes for software

            Data requirement determined by use (Scanning files use 1MB/hr during data acquisition.)

COM Ports                              UART 16550 AF supporting extended hardware buffering

SOFTWARE REQUIREMENTS

Operating System                   OS/2 Warp! Ver 3.0 or higher

SOFTWARE CAPACITIES

Run Time per Injection            655.35 minutes maximum;

                                               120 minutes maximum for scanning

Injections per Sequence         999

Calibration Levels                    Up to 15

Components                           750 identified peaks or 1500 unidentified

Integration                                3 signals selectabie for integration

Internal Standards                   Up to 3

Sample Names                       16-character name with 65-character description

Method Names                        99 characters including path and fite extension

Component Names                 15 characters